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Today I'm going to share with you the Neubauer/Improved Neubauer Haemocytometer that will help you be successful in your exams. This is going to be a long and detailed lecture post about Haemocytometer ,
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So, why is this topic importance in exam point of view? Well, did you know! There are about many lakhs students will appear for the exams in India and from abroad every year ?
Yes, that’s a lot of competition! But don't worry, We are here to help you successful in your exam. So we wish you to successful in exams, in this website post all your doubts and problems are solved.
We Will Talk About These topics!
Welcome to our detailed lecture on the Neubauer/Improved Neubauer Hemocytometer, an essential tool used in the field of hematology for counting cells and determining cell concentration. In this lecture, we will talk about the structure, components, and usage of the Neubauer Hemocytometer, providing step-by-step explanations to help you understand its operation and significance in cell counting procedures.What is Hemocytometer ?
A hemocytometer, also known as a counting chamber, is a specialized laboratory device or instrument that used for the manual counting of cells, particularly blood cells, in a given volume of fluid.
It consists of a rectangular glass slide with one or more counting chambers etched into its surface. Each counting chamber contains a grid pattern of squares, which allows for precise counting and measurement of cells under a microscope. Hemocytometers are commonly used in hematology, cytology, and other fields of biomedical research to determine cell concentration, assess cell viability, and monitor cell growth.
Structure of the Neubauer Hemocytometer:
1. Main Body:
The Neubauer Hemocytometer consists of a rectangular glass slide with precise dimensions and a thick, transparent cover slip. The slide is engraved with a grid pattern, which facilitates accurate cell counting and measurement.
2. Counting Chambers:
The central portion of the slide contains two counting chambers, each divided into a grid of tiny squares. Each counting chamber has dimensions of 1 mm x 1 mm, with a depth of 0.1 mm.
3. Grid Pattern:
The grid pattern on the counting chambers consists of nine large squares, each measuring 1 mm x 1 mm. Each large square is further subdivided into 16 smaller squares, creating a total of 400 smaller squares within the counting chamber.
4. Depth Markings:
The depth of the counting chambers is critical for determining the volume of sample analyzed and calculating cell concentration. Depth markings on the sides of the counting chambers indicate the thickness of the chamber, typically 0.1 mm.
5. Cover Slip:
The thick, optically clear cover slip is placed over the counting chambers to create a chamber of known depth and provide a flat surface for viewing and counting cells under a microscope.
Procedure of the Neubauer Hemocytometer :
1. Preparation of Sample:
Prior to cell counting, the sample (e.g., blood, cell suspension) must be properly diluted to ensure that the cell concentration falls within the counting range of the hemocytometer. Dilution factors are calculated based on the expected cell concentration and the desired volume to be counted.
2. Loading of Sample:
A small volume of the diluted sample (typically 10 μL) is carefully pipetted onto the edge of the cover slip, allowing the sample to be drawn into the counting chamber by capillary action. Excess sample is removed by blotting with absorbent paper.
3. Microscopic Examination:
The loaded hemocytometer slide is placed under a light microscope equipped with a calibrated ocular lens (typically 10x magnification). The counting chambers are positioned within the field of view, ensuring that the grid pattern is clearly visible.
4. Cell Counting:
Cells within the grid squares of the counting chambers are counted manually using a tally counter or by using a digital counting device. Only cells that are entirely within the boundaries of the square or touching the top and right lines are included in the count.
5. Calculation of Cell Concentration:
The cell concentration is calculated based on the number of cells counted, the volume of sample analyzed, and the dilution factor used. The formula for cell concentration is:
Cell concentration (cells/mL) = (Number of cells counted / Number of squares counted) × (1 / Volume of sample counted) × (1 / Dilution factor)
Advantages and Applications:
1. Accuracy:
The Neubauer chamber provides accurate cell counts and precise measurements of cell concentration, making it indispensable in various fields such as hematology, microbiology, and cell biology.
2. Versatility:
The hemocytometer can be used to count a wide range of cell types, including blood cells, bacteria, yeast, and sperm cells, making it a versatile tool for research and clinical diagnostics.
3. Cost-Effectiveness:
Compared to automated cell counting methods, the Neubauer Hemocytometer offers a cost-effective solution for laboratories with limited resources, providing reliable results with minimal equipment.
4. Quality Control:
The hemocytometer is used in quality control procedures to monitor cell viability, proliferation, and density in cell culture experiments, ensuring consistency and reproducibility of experimental results.
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Conclusion and Thank You:
In conclusion, the Neubauer/Improved Neubauer Hemocytometer is a valuable instrument for cell counting and concentration determination in various scientific disciplines. Its precise structure, coupled with meticulous sample preparation and microscopic examination, enables accurate and reliable cell counting results. As we continue to advance in the field of cell biology and laboratory techniques, let us recognize the importance of the Neubauer Hemocytometer in facilitating research, diagnosis, and quality control practices.
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