What is the role of cell culture in laboratory research?

Cell culture is the removal of cells either from plants or animals and cultivated in an artificial environment under controlled conditions for biological research. It is an invaluable tool to understand physiological processes, mechanisms of underlying diseases, effects of drugs and toxic compounds, and drug screening and development. Cell culture is also used in vaccine research, protein therapeutics, and cancer research. It is a meticulous process and one that requires a lot of diligence, skill, and experience.

Cell culture processes and procedures

The processes and procedures of cell culture depend on the cell type and application. There are two techniques of obtaining cells, from the cell bank or by isolating cells from tissue.

From Cell Bank

The cell culture procedure followed for cells obtained from the cell bank include:

Thawing – the cryopreserved cells are thawed in a 37°C water bath or a melting apparatus. Before the ice almost melts, the medium is added to dilute the cryoprotectant liquid, the cells are precipitated. The supernatant is removed, and a fresh medium preheated to 37°C is added. The cells are then resuspended using a pipette.

Cell Seeding – the desired cell seeding density is achieved by measuring cell numbers and dilute the cell suspension accordingly.

Cell Observation – after cell seeding, the cells are observed in the vessel with an optical microscope to check the viable cells, cells are evenly distributed in the vessel, presence of foreign objects, and check cell morphology. The cell culture vessel is then placed in a humidified CO2 incubator at 37°C to start culturing.

From Donor Tissue

When cells obtained from tissue, the procedure followed is:

Cell Isolation – the cells are isolated from the tissue using two methods, explant culture, and enzymatic method. In the enzymatic method, proteolytic enzyme solution is used, which is then diluted and then proceed to steps of cell seeding and cell observation to prepare cell culture.

Basic equipment and reagents required in this application

There are several equipment and reagents used in cell culture, these include:

Laminar airflow, incubator, pipettes, burettes, bottle top dispensers, inverted microscope, water bath, centrifuge, cell culture vessels, consumables, and many more.

The reagents are complete medium, buffered solutions, detaching agents, cryoprotective agents, and distilled water.

Benefits of using liquid handling instruments in cell culture

Cell culture involves meticulous liquid handling, which can be a tiresome and time-consuming procedure if not done properly. The slightest error can result in contamination and damage the cell culture. The liquid handling instruments satisfy the prerequisites as they are designed to improve liquid handling accuracy and precision, increase efficiency and avoid any chance of contamination.

The liquid handling instruments used in the cell culture process are pipettes, burettes, and bottle top dispensers. Serological pipettes are used to resuspend the cells. The burettes are used to dispense the accurate amount of reagent to detach, buffer, or isolate the cells. Bottle top dispensers are used in the sample preparation process to provide convenient dispensing of reagents. They significantly enhance productivity and prevent hazards.

Conclusion

The role of liquid handling equipment is essential in cell culture as they provide accuracy, precision and convenient sample preparation. The equipment minimizes contamination, reduces inconsistencies in sample yield, and helps prepare uniform quantities.

Microlit is a trusted manufacturer in providing high-quality liquid handling instruments for cell culture as well as other applications. To learn more about our products, please email us at info-usa@microlit.com.

Single-stage and Double-stage Micropipettes

Used in laboratories across industries like pharmaceuticals, F&B, environmental monitoring, life sciences, academics and more, pipettes are an essential tool and have been one since science has started to evolve. They are used to accurately measure and transfer a given quantity of a reagent from one vessel to another. A micropipette is capable of transferring volumes in the microliter range.

Almost all micropipettes operate on the same mechanism: Press the plunger with your thumb. When released, the reagent is drawn into a disposable plastic tip. When the plunger is pressed again, the reagent is dispensed.

In this blog, we’ll briefly discuss Microlit Lilpets, understand what single-stage and double-stage micropipettes are, how they’re different and the significance of that distinction.

What are Lilpets?

Microlit Lilpet, a miniature fixed-volume micropipette, is a cost-effective high precision liquid handling instrument with an ergonomic and intuitive design. It helps in performing sensitive operations with acute reliability in practical lab environments.

Some of the major fields where this micropipette is widely used include:

In schools for research and experiment purposes.
In mobile laboratories for soil testing and water testing, they enable one to easily go out and collect samples directly from the field.
In Rapid Testing Diagnostic Kits.

What are single-stage and double-stage micropipettes?

Most micropipettes have two ‘stops’ on their plunger. Before you can use the micropipette, it is advisable to test the plunger a bit to feel these two different stops. The first stop is the stop that dispenses a certain amount of reagent, but not in its entirety The second stop is an extra ‘push’ that is sometimes needed to remove the remaining reagent from the tip. So essentially, the two steps are a way to reduce any residual solution in the micropipette and get dependable results.

When it comes to Microlit Lilpets, there is an option to choose from single-stage and double-stage. They’re available in both configurations. The single-stage and double-stage Lilpets are respectively supported by single and double-column strokes.

In the single-stage Lilpet, only the first stop is depressed, which is the lowest position to draw and dispense the reagent. It is designed for users who may not be aware of the right pipetting technique or are using it for primitive tests or practice.
In double-stage Lilpet, the plunger is depressed to dispense the reagent up till the first step, and then the plunger is depressed further to dispose of the last drop. It facilitates reverse pipetting and reduces the loss of reagent dispensed, thereby enhancing the accuracy of the pipette. It is, therefore, quite suitable for test kits.

Even though two-stage micropipettes are more accurate when it comes to dispensing reagents, both the single-stage and double-stage micropipettes have very specific uses. Therefore, one must be very clear with their requirements while choosing between single-stage and double-stage micropipettes. Microlit has mastered both these pipettes that are ergonomically designed, aesthetically presentable and provide accurate and precise measurement values.

To get more information on different kinds of pipettes and purchase them, visit www.microlit.us or drop a mail at info-usa@microlit.com.

Polymerase Chain Reaction and its Applications

As the world continues to cope with the COVID-19 pandemic, there are certain terms that scientists have been using repeatedly and have now become a part of our everyday vocabulary. The most widely used is the RT-PCR test.

PCR is actually a widely used, simple yet very useful procedure in molecular biology. First discovered in 1983, the PCR amplifies or makes millions of copies of a particular section of DNA sample so that it can be studied in detail. This is a valuable technique used for DNA fingerprinting, diagnosis of genetic disorder and other molecular and genetic analyses.

The entire process is carried out in a controlled environment within a laboratory in a machine called a thermocycler. The process involves a continuous change in temperature known as thermal cycling to allow DNA denaturing and synthesis. In this blog, we will be discussing the significance and the various applications of the Polymerase Chain Reaction (PCR).

RT-PCR – A Household Name in Recent Times

As the COVID-19 disease is spreading across the world, the real-time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) test is playing a crucial role in its detection. The RT-PCR test is one of the fastest and the most accurate laboratory methods for detecting, tracking and studying the virus.

The PCR Process

The PCR test involves the following three steps. It might take a few hours or even less than an hour, with the help of a high-tech machine, to complete.

Image reference: https://www.yourgenome.org/

Steps involved in Polymerase Chain Reaction (PCR)

Step 1: Denaturation

The first step in the PCR test is to heat the double helix of the DNA to approximately 92°C. This high heat separates the 2 polynucleotide strands by breaking the hydrogen bonds of the DNA. This process is known as denaturation.

Step 2: Annealing

In the next step, the temperature is lowered to 50°C – 65°C. During this step, nucleotides and primers are added, and the primers bind to their targeted DNA sequences, initiating polymerisation.

Step 3: Extension

To achieve extension, the temperature of the reaction mixture is raised to 72°C. In this step, Taq DNA polymerase, an enzyme taken from the Thermus aquaticus, joins the free DNA nucleotides together. The end result that we get is two double-stranded DNA, one original and the other a newly made strand.

These 3 steps of thermal cycling are repeated 20-40 times to get a billion or more copies of the DNA. This process takes around 2-3 hours to complete.

Application of PCR

Diagnosis of Infectious Diseases

PCR, with its specificity, sensitivity and high speed of amplification has championed the identification of infectious diseases. It also allows the identification of non-cultivable or slow-growing microorganisms. The basis of this diagnostic is the detection of infectious bacteria or viruses. PCR helps in detecting diseases like human immunodeficiency virus (HIV), tuberculosis (TB).

Forensic Analysis

Genetic fingerprinting has seen widespread application in forensics. DNA profiling is used by forensic scientists for DNA typing, genetic fingerprinting, DNA testing, to identify someone based on their DNA. This technology is widely used by criminal investigators as it can identify one person from a million others.

DNA Cloning

A molecular biology technique, DNA cloning enables the lab researchers to make identical copies of a piece of DNA. In a typical cloning process, the fragment of interest is inserted into a larger genome, amplifying the DNA. With slight alterations to the PCR protocol, genes can be mutated.

Role of Liquid Handling Instruments in PCR

Though most of the PCR process is automated and easy-to-conduct, it is the setting up which is highly time-consuming. The PCR setup involves numerous pipetting steps that can act as a bottleneck and also increases the chances of human error as well as contamination.

Given the current conditions, for instance, the whole process and getting the correct results is crucial. During the setup, microlitre or even sub-microlitre volumes of a reagent are transferred, automatically increasing the chances of error resulting in inaccurate results.

Then there is speed. To carry out such a tedious task it is imperative that no step should be hurried. Therefore, the setup takes time. It will be highly beneficial if the speed of the process can be increased, helping in carrying out more tests at the same time.

To curb these challenges, the answer is automation.

Using sophisticated, automated liquid handling instruments during the setup of PCR will be highly beneficial for lab users. With a fully automated PCR, the certainty and reliability of the test will increase while saving time. Automation also means minimal human contact eliminating the chances of contamination and degradation of the sample.

The Microlit Micropipettes

Micropipettes are sophisticated, high precision instruments designed to accurately measure and transfer volumes in the microlitre range. One of the most widely used scientific instruments, micropipettes are widely trusted to be accurate and hassle-free. As discussed above, they are used during the sample preparation stage of a PCR process. The available variants cater to the requirements of the experiment to be conducted.

Single Channel Variable Volume

A high precision micropipette, available in unique volume ranges, prominently used for Molecular Biology, Microbiology, Immunology, Biochemistry, Genetics. Explore Microlit Single Channel Variable Volume Micropipettes.

Single Channel Fixed Volume

A high precision micropipette, available in 15 unique volumes, recommended for Clinical Diagnostics, Control Analysis. Explore Microlit Single Channel Fixed Volume Micropipettes.

Multichannel (8-Channel and 12-Channel)

High precision micropipettes, available in 6 unique volume ranges, recommended for ELISA (Diagnostic Test), Molecular Screening, Kinetic Studies, DNA Amplification. Explore Microlit Single Channel Variable Volume Micropipettes.

When it comes to accuracy and precision, Microlit has earned a reputation of being a company that ergonomically designs its products ensuring the customer gets what s/he needs. With our range of innovative liquid handling instruments, we focus on blending features and functionality that facilitate a remarkable user experience in the lab.

If you routinely conduct PCR and related tests in your lab, ensure greater precision and accuracy with Microlit Micropipettes and Filter Tips. Get in touch with us at info-us@microlit.com.

E-Burette Product Guide

What is a Burette?

Burette is an essential laboratory instrument, commonly used in the titration process for quantitative analysis in many industrial chemical tests where solutions of known concentration are used to find the concentration of unknown solutions.

Burette Uses in Laboratory

Burettes are vital in laboratories for precise liquid measurement and dispensing. Commonly used in laboratory titrations, they determine unknown solution concentrations accurately. Their versatility extends to various applications, including acid-base and redox reactions. Burette use is crucial in quantitative chemical analysis, making them indispensable in scientific research and educational settings. Uses of burette also encompass base burette laboratory apparatus uses, highlighting their integral role in diverse experimental procedures.

History of Burette

The burette, a cornerstone in chemical analysis, traces its origins to late 18th-century France. Its development is credited to the French chemist Joseph Louis Gay-Lussac, who enhanced its design to enable precise titration. This groundbreaking innovation marked a significant milestone in analytical chemistry, allowing scientists to measure and dispense liquids with exceptional accuracy. Gay-Lussac’s improvements transformed the burette into a reliable tool for quantitative experiments, revolutionizing laboratory practices. Over time, the burette evolved further, incorporating advanced materials and features to enhance precision and ease of use. Today, it remains an essential instrument in laboratories worldwide, symbolizing the progress of scientific innovation.”

Before learning more about a Burette function in laboratory, let’s get to understand the titration process in detail.

What is a Titration?

Titration is a method commonly used in a laboratory which uses a solution of a known concentration to analyse and determine the known concentration of another solution. Typically, the titrant (the known solution) is added from a burette to the known quantity of the analyte (the unknown solution) until the reaction is complete. Often, an indicator is used to usually signal the end of the reaction, the endpoint.

Types of Titration?

There are several types of titration with different goals and processes. However, the most common types of titration in quantitative chemical analysis are redox titration and acid-base titration.

These four common titrations are:

Acid-base titrations
Redox titrations
Precipitation titrations
Complexometric titrations

1. Acid-base Titration:

Acid-base titration is a chemical testing process used to determine the unknown concentration of an acid or base by neutralizing the other acid or base of known concentration. Acid-base titration can also be used to measure the purity of chemicals.

HA + BOH → BA + H2O

Acid + Alkali → Salt + Water

Or (H+) + (OH–) → H2O

Acid-base titration is based on the reaction that neutralization occurs between an base or an acidic and an analyte. In this kind of titration, a reagent is mixed with the sample solution until it reaches the required pH level. This type of titration mainly depends on the track change in the pH or pH meter.

2. Redox Titration:

Redox titration is a laboratory method of determining the concentration of a given analysis by creating a redox reaction between the titration and the analyte. Sometimes, This type of titration needs the use of a potentiometer or redox indicator. It is also known as Oxidation-reduction reaction.

Redox titration involves a transfer of electrons between the used Analyte and the titrant. These kind of titrations are named after the reagent that is used in are as follows:

Permanganate Titrations
Dichromate Titrations
Iodimetric and Iodometric Titrations

Permanganate Titration

In this, the potassium permanganate is used as an oxidizing agent maintained with the use of dilute sulphuric acid. The solution remains colourless before the endpoint. The equation is:

2KMnO4 + 3H2SO4 → K2SO4 + 2MnSO4 + 3H2 + 5O

Or MnO4– + 8H + 5e → Mn2++ 4H2O

Dichromate Titration

In this titration, potassium dichromate is used as an oxidizing agent. The medium is maintained acidic by use of dilute sulphuric acid. The solution of potassium dichromate can be used in titration directly. The equation is:

K2Cr2O7 + 4H2SO4 → K2Cr2(SO4) + 4H2O + 3[O]

Or Cr2O27- + 14H + 6e → 2 Cr3+ + 7H2O

Iodimetric and iodometric Titration

The reduction of free iodine to iodide ions and oxidation of iodide ions to free occurs in Iodimetric and iodometric Titrations.

l2 + 2e → 2l–……………. (reduction process)

2l– + 2e → 2e ……………. (oxidation process)

The solution is used as an indicator. Free iodine is used in the iodometric titration, while in the iodometric titration an oxidation agent is used to react the process to liberate free iodine.

3. Precipitation Titration:

Precipitation titration is based on insoluble precipitation where two reacting substances are brought into contact, called precipitation titration. The titrant reacts with the analyte and forms an insoluble material. The titration continues until the analysis is completely consumed. When the titrant is excess it reacts with the indicator and signals to end the titration process.

For example, To determine the concentration of chloride ion in a certain solution, we can titrate this solution with a silver nitrate solution which is of known concentration. The reaction occurs as follows:

Ag+(aq) + Cl–(aq)→ AgCl(s)

As a white precipitate, AgCl can be seen to settle at the bottom of the reservoir during titration. The amount of silver ion used for the equivalence point is basically equal to the amount of chloride ion present.

4. Complexometric titration:

The complexometric titration is where an undivided complex is formed at an equivalence point. This is greater than the precipitation titrations, and there will be no error due to co-precipitation.

Hg2+ + 2SCN– → Hg(SCN)2

Ag+ + 2CN– → [Ag(CN)2]–

Ethylenediaminetetraacetic acid (EDTA) is an important solution that forms complexes with metals.

Detecting the end-point of a titration

End-point is a physical change of solution that indicates the completion of titration. It is used as a signal for testers to stop adding more titrant to the analyte. Usually, it is indicated by any form of a visual indicator like color change, a visible precipitate, or a change on an electronic readout.

Performing a routine titration to determine the concentration of a analyte

The following steps use a conventional glass burette to do the titration. The same experiment can also be performed with the help of an electronic burette, a better option that minimizes the risk posed by multiple sources of error that may occur with conventional glass burette:

Place a known volume of the solution of unknown concentration (the analyte) in a flask and fill the solution of known concentration (the titrant) in the burette.
Slowly open the burette tap and allow the titrant to come down in the analyte. Determine the correct titration speed – too slow and the titration duration is unnecessarily prolonged; too fast, and the equivalence point is exceeded.
Keep track of the color of the analyte – completion of titration is usually indicated by color change.

Finally, if the titrant and analyte are in the 1: 1 mole ratio, use the following equation to determine the concentration of the analyte.

M_titrant x V_titrant = M_analyte x V_analyte

where,

M_titrant = Molarity (M) of the titrant,

V_titrant = Volume (V) of the titrant,

M_analyte = Molarity (M) of the analyte, and,

V_analyte = Volume (V) of the analyte.

Note: Molarity is the concentration of a solution expressed as the number of moles of solute per liter of solution.

Application of the Titration

Titration is actually referred to as volumetric analysis. It is a technique on which many industries rely on the development and analysis of major chemical compounds. Here are some notable uses in titration in various industries that affect many aspects of society:

Pharmaceutical Industry: A particular amount of chemicals is prescribed for the development of drugs. This measurement of the amount of chemicals is finalized by the process of titration. Various titration procedures such as purity analysis, content analysis, precipitation titrations and pH-stat titration are used in the pharmaceutical industry.

Wine Industry: Wine makers use titration during the winemaking process. This allows them to check the quality of the raw materials used for production and to bring out the flavor of the wine. The titration also helps to test the batch during fermentation, and is used for final quality control.

Automotive Industry: The automotive industry uses the titration process to produce biodiesel oil. A special pH level is required to ensure the functionality of biodiesel oil. For this purpose, titration is useful. Manufacturers measure the pH level of biodiesel and then determine the amount of base used to achieve the desired pH level.

Food & Beverage Industry: The Food & Beverage industry primarily uses titration techniques for two purposes – regulatory requirements and quality control. Every producer should test and measure the quantity of different minerals in the food & beverage production. The presence of these minerals is determined by precipitation and complexometric titration.

Manufacturers use acid-base titration for quality control. It is important to control the taste, texture, nutrition, consistency and appearance of produced food.

Cosmetic Industry: To ensure quality production of cosmetics, manufacturers need to add appropriate concentration and quantity of chemicals. The titration process helps manufacturers determine the most suitable base for cosmetics, for example, coconut oil is used as a base in many products.

Burette Uses in Laboratory: Burette is a laboratory apparatus commonly used to dispense and measure variable amounts of liquid or sometimes gas within chemical and industrial testing specially for the titration process in volumetric analysis.

Burettes can be specified according to their volume, resolution, and accuracy of dispensing. Smaller diameter tubes result in lower volume and increased resolution. Digital burettes can achieve better accuracy and are less dependent on the skill of the technician.

The first burette was created by chemists in the 19th century. It consisted of a simple glass tube with valves and later added graduations. This basic design is still widely used in glass burette. This should be carefully observed at eye level, ensuring that the liquid is free of bubbles and the reading is taken at the bottom of the meniscus. The delivered volume is the difference between the initial and final readings of reagent.

Types of Burette

There are three types of burette:

In laboratories, types of burette include Volumetric, Piston/Digital, and Electronic. Each type of burette serves specific functions, making them essential for precise liquid measurements in various experiments. Understanding these types of burette is crucial for accurate laboratory work.

1. Volumetric Burette

A volumetric burette is made of glass or plastic material, with a volume scale printed on the burette wall. It has a stopcock at the end of the instrument, and a valve to control the flow of liquid. The barrel of a stopcock is made of glass or plastic. Burettes are usually fixed with a clamp in an iron stand. Normally, volumetric burette can be classified as:

Liquid burette
Gas burette

On a liquid burette, it has a long graduated glass tube with a stopcock at the bottom to control the flow of liquid. Gravity causes fluid to flow when the valve is opened. The exact amount of liquid can be determined by reading the volume marked on the glass tube.

In a gas burette, the stopcock is given at the upper side of the burette. The tube of the burette is filled with a fluid, such as water, mercury or other kind of liquid and is attached to a reservoir of fluid at the bottom of the tube. The gas is collected by displacing the fluid from the burette, and the volume of the gas is measured by the volume of the displaced fluid.

2. Piston or Digital Burette

A piston or digital burette is based on a syringe design wherein the barrel and plunger can be made of glass or plastic. For alkali solutions, the barrel and plunger may be made of polyethylene or another resistant plastic material. The barrel is placed in a fixed position and the plunger is moved incrementally by rotating the wheel by hand or by means of a step motor.

The volume is shown on a digital display given at the top of the burette. A high-precision syringe can be used to deliver a very precise amount of liquid. Motorized digital burette can be controlled by a computer.

3. Electronic Burette

The Electronic Burette or E-Burette has an electronic display, electronic control panel and motor controlled piston movements that eliminates all problems of the digital burette which dispense manually. The accuracy of electronic burette is higher than a digital burette due to motor controlled dispensing. This motorized system eliminates the human errors from the titration/dispensing process that makes the titration more precise.

Components of a Burette

Components of Volumetric Burette:

Volumetric or glass burette consists of a graduated glass tube with a stopcock at one end. The precise liquid dispensed can be determined by volume marking given on the glass tube surface. A Tip is given at the other end of the glass burette which is used to dispense liquid precisely. The Stopcock helps to control the flow of liquid by rotating it clockwise or anti-clockwise.

Components of Electronic Burette:

E-Burette is an electronic device that delivers accurate and convenient titration while reducing the risk of accidental chemical contact. It includes some advanced components that reduce manual operation with high work efficiency. Some major components of E-burette are:

Recirculation Valve – It prevents the loss of reagents during purging by re-directing them into the mounted bottle.

Delivery Tube – Delivery tube provides the facility to deliver the reagent beyond the reservoir.

Inspection Window – The transparent acrylic window can be replaced with an amber colored window for the use of chemicals. These chemicals such as iodine, potassium permanganate, and silver nitrate solutions are sensitive and can react in light. To protect these chemicals from such reactions, an amber-colored window is used that can be easily changed.

Delivery nozzle – Delivery nozzle provides a high degree of flexibility in both the horizontal and vertical directions, facilitating easy dispensing according to different laboratory conditions.

TFT Screen – A user-friendly TFT touchscreen guides quick and accurate titrations and displays numeric data up to the second decimal. It is easily operated with Stylus.

Telescopic tube – The telescopic tube is used to draw up the liquid from the reagent bottle to e-burette. It can be adjusted in length according to a variety of bottle sizes.

Adapter – E-burette comes with different standard sizes of adapter that comfortably fit most laboratory reagent bottles. The standard sizes of adapters are 28mm, 32mm, 38mm, 40mm and 45mm.

Stylus – It is a sleek hand-held instrument like pen used to operate the control panel with ease.

Control Panel – E-Burette comes with a powerful control panel that is used to perform functions like automatically re-fill without changing the reading, perform a zero reset (via. tare). switch the instrument off etc.

Middle Housing – It consists of a piston mechanism inside the burette.

Components of Digital Burette:

Digital burette is different from e-burette, although many components have the same functionality. Recirculation Valve, Delivery Tube, Inspection Window, Telescopic tube and Adapters have the same functionality like Electronic Burette. The digital burette only has an electrical display but the piston movement is manual which can be operated manually using a wheel.

A digital display allows the user to note the volume of the liquid dispensed out of the burette. This feature eliminates meniscus reading errors. A hand wheel gives you the facility to set the amount of dispensed liquid by rotating it clockwise or anti-clockwise.

Various buttons including ON/OFF button, CLEAR/Selection button and Pause Button are available at upper housing of the digital burette that helps to control the dispensing process.

Technical Specifications of Burette

An Electronic Burette comes in different sizes and the most popular volume ranges are 10ml, 25ml and 50ml in capacity. The technical specification of these burettes are listed below along with their permissible error limits as specified in the DIN EN ISO 8655-3.

Capacity (ml)	Increment (ml)	Accuracy (± %)	Accuracy (±ml)	CV (± %)	CV (±ml)
10	0.005	0.1	0.01	0.1	0.01
25	0.01	0.07	0.0175	0.05	0.0125
50	0.01	0.05	0.025	0.05	0.025

How to use a Burette?

The main steps to use a Electronic burette are given below:

Connecting the batteries

Open the battery panel at the back of the control panel.
Connect the battery connector to the panel wire by inserting the battery connector into the panel, and then close the panel.
Press the ON/OFF button. The screen will display “Welcome”.

Setting Up

Adjust the length of telescopic tube according to reservoir bottle length
Fix the telescopic tube at the bottom of the burette. Fix it from the wider end.
Choose the correct adapter according to the reservoir bottle and fix it by screwing it in a clockwise direction.
Mount the instrument on a reservoir bottle by screwing it with gentle hand torque.

Connect the cable from the instrument housing to the port at the back of the control panel.
If the charging is required, insert the charging pin at the back of the control panel and connect the power cord. Battery indicator will show the charging.
Now E-burette is now ready to be switched on.

Operating the Instrument

Switch on the instrument using the ON/OFF switch at the back of the control panel.
When it boots, the instrument will warn the user to switch the knob to recirculation mode.
After turning the knob to recirculation mode, click the tick mark on the control panel screen when ready.
Instrument will reset automatically, now it is ready to use.

Before starting using the digital burette, it is recommended to ensure bubble free dispensing:

Click the purging mode from the home screen on the control panel.
The screen on the control panel will warn the user to turn the knob to recirculation mode.
Turn the knob to recirculation mode and click the tick mark on the control panel screen when ready.
The digital burette will purge automatically. If the instrument is still not properly purged, repeat the purging process again till no large air bubble is visible below the piston.

E-burette is commonly used in the titration process to determine the concentration of an unknown solution. Typically, titrant (known solution) is added from a burette to a known volume of analyte (unknown solution) until the reaction is complete. An indicator is used to find the endpoint of the titration.

Once the E-burette is properly setup, follow the steps below to perform the titration process.

Remove the cap from the discharge tube.
Click on the titration mode on the home screen. The screen on the control panel will warn the user to turn the knob to titration mode.
Turn the knob clockwise to set at titration mode and click the tick mark on the screen of the control panel when ready.
Place the discharge tube orifice against the inner wall of a suitably receiving vessel.
Now fill the barrel from the titration screen by clicking the fill button. By pressing the stop button, the filling volume can be stopped as and when you need to.
Microlit E-burette offers the free dispensing speeds are provided, including dropwise dispensing that allows the user to get accurate end point of the titration:

Burette Model	Mode	Speed
50 ml	Fast	3.8ml/sec
	Medium	1.3ml/sec
	Dropwise	10ul/click
25 ml	Fast	2.1ml/sec
	Medium	0.7ml/sec
	Dropwise	10ul/click
10 ml	Fast	1.2ml/sec
	Medium	0.2ml/sec
	Dropwise	5ul/click

Ths user interface is very responsive and the dispensing starts by clicking the appropriate button on the control panel and stops immediately when the touch is removed.
Press the TARE button to reset to Zero and repeat the process.

Difference between Burette and Pipette

Burette and pipette are both analytical tools. However, a significant difference between the burette and the pipette is in the release mechanism. The burette has an open top and small exit point at the bottom of the tube, which closes with a stopcock. A tester pours liquid into the burette from the top until the required volume reaches the marker. However a digital burette has an automated system to draw up the exact amount of liquid from the bottle.

A pipette requires a suction device at the top end, which changes the pressure inside the tube to pull the liquid upward through the tapered end from a container of liquid. As long as the suction pressure remains on the pipette, the liquid remains inside the tube. When the suction is removed, the liquid falls out because there is no mechanical locking system in the lower end of the tube like a burette.

Usually, Burettes are larger than pipette with higher capacity of liquid to dispense. That’s why, the accuracy of burette in releasing small volumes is lesser than pipette. Also, most of the advanced burettes have completely automated systems while pipettes are still hand operated to dispense desired amount of liquid.

Burette Vs Pipette

	Burette	Pipette
Definition	A graduated glass tube with a stopcock at one end, used to deliver a known amount of a liquid, specially for titrations.	A pipette is a common yet an essential laboratory tool, used to accurately and precisely transfer desired volumes of liquid in the microliter range.
Releasing Mechanism	Have a stopcock at the end to control the release of the liquid.	Has a dropper like system that releases desired amount of liquid by lessening vacuum.
Size	Usually larger than a pipette.	Usually smaller than a burette.
Accuracy	Less accurate in dispensing small amount of liquid.	More accurate in dispensing small amount of liquid.
Operating	Most of the advanced burettes are automated or very less manual.	Completely hand-operated.

Difference between Burette and Bottle Top Dispenser

The basic difference between a Burette and a Bottle Top Dispenser is its use and accuracy. Burette is used exclusively for the titration process, where the Bottletop Dispenser provides only functionality of the Liquid dispensing. Due to the large amount of liquid handling with the bottle top dispenser, the accuracy of BTD is much lower than that of Burette.

Another major difference between burette and BTD is its functionality. Burette gives you the functionality to dispense the liquid in different modes where you can also identify the precise volume of dispensed liquid. BTD has no feature to identify the dispensed volume of liquid that makes it useless for titration. BTD can transfer the precise amount of liquid in a single dispense circle.

Cleaning, Maintenance and Storage of a Burette

The maintenance of electronic burette is an important routine in any chemical laboratory. Adopting a proper maintenance schedule can reduce the cost of purchasing new equipment. Cleaning the e-burette takes time and practice, otherwise the equipment may deteriorate. Therefore it is necessary to handle the burette carefully while cleaning.

The instrument must be cleaned in the following situations to ensure the correct operations and test results:

Immediately, if the piston become harder than normal use
Before changing the reagent
Prior to long term storage
Prior to dismantling the instrument
Regularly when using crystallizing liquids
If the liquid has accumulated in the screw cap of the titration tube

Note: The piston cylinder, valves, telescoping filling tube and titration tube may contain the reagent. The necessary safety steps must be taken before cleaning the instrument.

Standard Cleaning of Burette:

Whenever cleaning is required, run the burette under distilled water.

Fill the barrel completely with distilled water and press the fast dispense button to dispense the water completely.
Place a suitable receiving vessel below the opening of the discharge tube, and rinse the instrument several times by completely filling and emptying it.
Set the valve to recirculation and rinse the instrument several times by completely filling and emptying it.
This process can optionally be repeated with a suitable cleaning agent.
Unscrew the chuck nut by rotating it in anticlockwise direction and pull out the delivery pipe. Clean the delivery pipe with deionized water and assemble it again.
Next, rinse the burette again with deionized water.

Intensive Cleaning of Burette:

The dispensing cylinder, valve, telescoping filling tube and titrating tube are filled with reagent. For this reason, always perform a standard cleaning before dismantling the instrument.

In order to avoid confusion about the components, do not dismantle more than one instrument at a time. A calibration, and any necessary adjustment, must be carried out after dismantling or replacement of a piston/cylinder assembly.

Note: Use rubbing alcohol on a cloth or cotton to clean the external body.

Proper storage of digital burette is as important as cleaning and calibrating the instrument. The digital burette and its all accessories must be stored in a clean, cool and dry place. The storage temperature should be ranging from –20 °C to +50 °C (from –4 °F to 122 °F) is advised with relative humidity between 5% and 95%. Another point to be remembered is that the instrument should be stored in an upright position as recommended by the manufacturer. .

Calibration of a Burette

A gravimetric testing based recalibration of the burette is recommended every 6-12 months depending on the following factors:

Frequency of use
Changes in environmental conditions
The viscosity of the media

For Microlit E-Burette, gravimetric volume testing according to DIN EN ISO 8655-3 should be performed as follows:

Click the settings icon on the bottom right of the control panel home screen.
From the settings menu, choose calibration.
Fill the nominal volume of double distilled deionized water by clicking the FILL button on the calibration screen of the operating panel.
Dispense the filled liquid by clicking on the DISPENSE button on the calibration screen of the control panel.
Measure the dispensed liquid on a balance and enter the value in mg using the provided electronic keyboard. Click the tick mark to proceed. CAL label will start appearing on the top of the screen.
Repeat the procedure until the nominal volume is achieved on the electronic balance.

Calculation of a Burette Reading

Calculation of accuracy (A %) and coefficient of variation (CV %) are calculated according to the formulas for statistical control.

When is burette calibration required?

In order to maintain the quality of titration results, the burette must be in good condition and properly calibrated. Generally, the interval at which a burette needs to be calibrated depends on several factors:

Frequency of burette use
Type of liquid dispensed by the burette
Long period of non-usage
Handling and care of burette
Titration applications that require superior accuracy also demand more frequent calibration

Regulations and standards published by organizations such as the FDA and ASTM International provides minimum requirements to ensure the quality of laboratory testing results. Regulations specify that all laboratory instruments used in sampling & productions including pipette/ burette must be routinely calibrated at suitable intervals.

In the same series, the Clinical and laboratory standards institute (CSLI) has provided the guidelines for pipette/ burette that specifies these instruments must be calibrated every 3 to 6 months. And a minimum of two volumes must be tested with 10 replicas at both nominal and lowest settings.

Establishing an appropriate calibration frequency minimizes the chances of incorrect liquid delivery in the laboratory and ensures traceability, accountability and confidence in the results.

Applications of a Burette

Burette is commonly used in the titration process for quantitative analysis in many industrial chemical testing where a solution of known concentration is used to find the concentration of an unknown solution. The process of titration is widely used in various industrial applications listed below:

Pharmaceutical industry: Purity analysis, Content analysis, Precipitation titration, pH-stat titration
Wine industry
Automotive industry
Food and Beverage industry
Cosmetic industry

Choosing the right Burette

Choosing the best burette for your application can be a challenging task as the liquid handling market has a variety of options. Before selecting a burette for your lab, it is necessary to consider a few questions:

Is the burette able to handle your typical volume range of liquid required for titration?
Does the burette provide different speeds for dispensing for proper titration?
Is it operated manually or automatically?
Is the accuracy and precision in reading offered by the burette in accordance with the ISO standards?

The above points ensure correct burette procurement that will work for a long time and improve titration activities in your laboratory.

Microlit E-burette is an advanced laboratory instrument designed keeping ergonomics and smooth operation in mind. Microlit E-burette provides sophisticated burette features and burette functionality such as touch screen enabled control panel with graphical user interface (GUI), motor-controlled piston movement and three dispensing speeds to perform accurate titrations.

3 Pre-set Dispensing Speeds

Microlit E-burette offers the 3 calibrated pre-set speeds that allow the user to work at the desired speed while avoiding manual error.
The dropwise dispensing option in the MicroLit E-burette allows the user to choose this mode when they are about to reach the end-point of the titration process. The dispense droplet can be as small as 5.0ul and record a highly accurate end-point of the titration.

TFT Touch Screen Control Panel

The TFT touchscreen of the control panel helps the user to perform quick, easy and accurate measurements, and displays numerical data up to two decimal points. The user needs a Stylus and keeps it pressed against the screen to perform dispensing and lift it to stop. The touchscreen feature also omits any parallax error which normally occurs in glass burettes when it is not viewed from a perpendicular position.

Motor Controlled Piston Movement

Dispensing is done manually in glass and digital burettes leading to inaccurate results. However the MicroLit E-burette has a motor controlled piston speed that allows laboratory personnel to work efficiently as it does not need to oversee the delivery process at every moment. 3 Pre-set speeds reduce human effort as well as errors which makes it the most efficient lab instrument for liquid handling.

Burette Troubleshooting

Problem	Possible Cause	Solution
Piston difficult to move	Piston assembly is contaminated or damaged by crystal deposits Deposition of dirt	Perform a cleaning Replace the piston assembly
Filling not possible	Filling valve stuck	Clean the filling valve. If the valve ball is stuck, use a 200ul plastic tip to loosen it
Air bubbles in the instrument	Instrument filled too quickly Purging not complete Filling tube loose or damaged Filling tube does not dip into the liquid Recirculation tube is not mounted or mounted improperly	Fill instrument slowly Purge the instrument again Fasten the telescoping filling tube firmly. If necessary, cut the tube off approx. 1 cm from the top of replace it Fill up the bottle, or correctly adjust the length of the telescoping filling tube Attach the recirculation tube. The opening must point outward toward the bottle wall
The volume delivered is smaller than that indicated	The instrument has not been completely primed Filling valve is blocked or damaged	Prime the instrument again Clean the filling valve, and if necessary replace the filling valve
The instrument doesn’t indicate any function	Internal error	Perform a restart

For more information about Microlit E-Burette, visit: Microlit E-Burette.

New Drug Development and the Role of Liquid Handling Instruments

The past few years have seen an upward rise in the number of novel diseases. It has put human life at risk and jeopardized the working of the world in several ways. More recently, COVID-19 caused by SARS-CoV-2 has disrupted our way of living.

New diseases make the development of new drugs extremely essential and important. It’s a meticulous task requiring a lot of diligence, skill and research. Let’s take a look at the process in more detail.

Developing new drugs

Shown above: Five key stages of new drug development

There are five key stages to developing a new drug:

The first one is the discovery and the development stage. Here, the groundwork for the new drug is laid out. Extensive and exhaustive research is done followed by several intensive tests of the molecular compounds.
The next step is Preclinical Research, where the new drug undergoes several lab tests. These tests are performed to gauge the safety and efficacy of the drug.
Once the results from the Preclinical Research are assessed to be favorable, the drug development process moves to the next step – The Clinical Research. Here, the drugs are tested on a random population and their effects observed. This is done to further strengthen the claim of safety and effectiveness.
Once a satisfactory amount of data has been collected through the preclinical and clinical research steps, the new drug being developed undergoes a Drug Review. The designated team thoroughly examines the data submitted by the company responsible for the development of the drug and decides whether the drug should be approved for usage or not.
Finally, even when the drug is approved for mass usage and is being sold in the market, it is still monitored by a designated team of professionals under Post-market Drug Safety Monitoring.

Benefits of using liquid handling instruments in new drug development

The process of drug development is a task requiring extreme precision. It is crucial to negate the possibility of error altogether. This can be achieved by using the right kind of instruments and apparatus.

Liquid handling equipment satisfies these prerequisites. The ones used for the development of a drug can vary from a basic pipet to a complex workstation. The more appropriate and effective liquid handling equipment for drug development include micropipettes, pipets, dispensers and burettes. These are vital because:

Liquid handling instruments offer high flexibility. They can be used in a variety of applications – pipetting, transferring, diluting, dosing, or dispensing – and can help in the transportation of cells, proteins, oligonucleotides or other biological components. This high flexibility is facilitated by the equipment’s throughput, capacity and dynamic volume range.
Previously, in the absence of the right technologies, liquid handling instruments were proving to be ineffective in bringing down the cost of quality control processes. However, advanced liquid handling instruments can easily facilitate the swift and repeatable assessment of the accuracy and precision of the volumes dispensed.
Owing to their sophisticated functionality and high precision capability, these instruments ensure that clinical research results are more reliable and accurate and false-positive* and false-negative* results are minimized.

*Significance of false-positive and false-negative results

False-positives: They are not fatal to the drug discovery process, but decrease efficiency, causing researchers to waste time and resources chasing dead ends. Reducing the number of false positives pursued through the drug discovery process can decrease the number of retests, minimise reagent loss and maximise staff and equipment utilisation.

False-negatives: Dispensing reagents in volumes inaccurate by even minuscule amounts, especially in complex serial dilution assays, can alter the concentration and prevent the identification of a reaction between compounds that are the foundation for a new drug. It is impossible to estimate the value of lost revenue from this type of error, but generally, this amount can sometimes run into billions of dollars.

Instrument-wise applications

1. Single Channel Micropipettes

They are used to dispense a fixed volume of a reagent in the microlitre range into a vial, test tube, petri dish or beaker. For instance, transferring samples related to cell culture, molecular biology, microbiology, immunology, genetics and more. Explore Microlit Single Channel Micropipettes.

2. Multi-channel Micropipettes

Dispensing fixed volume of a reagent in the microlitre range into assay plates, ELISA trays, 96 well plates – all this comes in the purview of a multi-channel micropipette. For example, they are used in ELISA (diagnostic test), molecular screening, kinetic studies, DNA amplification, etc. Explore Microlit Multi-channel Micropipettes.

3. Bottle Top Dispensers

They are essentially used in dispensing fixed volume of a reagent in the millilitre range into a beaker, flask or test tube. They are used for sampling and analysing Active Pharmaceutical Ingredients (APIs), impurities, excipients, etc. Explore Microlit Bottle Top Dispensers.

4. Burettes

Their major application is performing titration by adding the titrant (solution of known concentration) to the analyte (the second solution) contained in a titration flask. Burettes can be used with the most common titration solutions (max. 1 mol/l), such as sodium hydroxide, hydrochloric acid or iodine-potassium iodide solution. Explore Microlit E-Burette.

On a concluding note

The role of liquid handling instruments is not just important to the development of a new drug but also essential to the process. With the rapidly burgeoning pharmaceutical and related industries, their uses and significance are swiftly increasing as well.

Microlit has been a trusted and reliable partner to several top pharmaceutical companies, helping them make the world a healthier place, one drug at a time.

To know more about our products or buy them, please write to info-usa@microlit.com.