Observation experiments on immune-related cell morphology

Summary

All living things are made up of cells, only the size and shape of the cells vary from organism to organism. Some cells are visible to the human eye, such as the eggs of birds, the largest of which is nearly 10 centimeters in diameter (ostrich eggs).

Operation method

cellular observation

Materials and Instruments

Cell Smear
Ritter's stain Methanol
Microscope Slides Coverslips Droppers

Move

I. Observation of blood smears
1. Erythrocytes: light red, round cells without nucleus, because the red blood cells are biconcave, so the edge part of the staining is darker, the center is lighter, 7~8 microns in diameter.
2. Particulate white blood cells(1) Neutrophilic granular white blood cells: slightly larger than erythrocytes, the nucleus is stained purple and lobulated, which can be divided into 1~5 lobes, and between the lobes are associated with chromatin filaments, which are stained pink and filled with tiny uniformly sized granules stained purple-red. Diameter 10~12 microns.(2) Eosinophilic granular white blood cells: slightly larger than mesophilic white blood cells, nuclei stained purple, usually 2-lobed, cytoplasm filled with eosinophilic large round granules, stained bright red. The diameter is 10-15 micrometers.(3) Basophilic granular white blood cells: slightly smaller than eosinophilic white blood cells, cytoplasm with granules of different sizes stained purple, the number of granules is less than that of eosinophilic white blood cells, nucleus is 1~2 lobes stained light blue. Diameter 10~11 microns.
3. Granulocyte-free white blood cellsLymphocytes: both small and medium sized can be observed in the smear. Small lymphocytes are similar in size to red blood cells and are round. They contain dense nuclei stained dark purple. They are surrounded by only a thin layer of alkaliphilic cytoplasm stained light blue. Medium lymphocytes are larger, with broader layers of cells and round nuclei. 6-8 micrometers.Monocytes: largest, rounded cells. Cytoplasm stains gray-blue. The nucleus is kidney or horseshoe shaped and stains slightly lighter than the nucleus of lymphocytes. Diameter 14-20 micrometers.
II. Observation of mast cells
The cytosol is large and ovoid, and the cytoplasm is filled with coarse and homogeneous alkalophilic granules. It contains heparin, histamine and other substances. They are often distributed in groups around blood vessels.
III. Observation of plasma cells
The cells were round or oval, with abundant cytoplasm and alkalinity. The nucleus is round, deeply colored, mostly on one side of the cell, and the chromatin nuclear membrane is distributed in wheels. Normal tissue plasma cells are few, and increase in chronic inflammation. Plasma cells synthesize and secrete antibodies, which are important for immunity.
IV. Macrophages

Also known as histiocytes, the cell morphology is irregular. Often extend short and blunt protrusions, have a strong phagocytic ability.

Common Problems

Ritter's stain:

1. Configuration of the staining solution

Dissolve 0.1 g of Ritter's dye in 60 ml of methanol and filter. Store in a brown bottle. (When configuring, put the Ritter's Dye into a mortar and pestle first and add methanol drop by drop while researching to make the dye solution better.)

2. Ritter's staining method:

Take blood from mouse bone artery and smear the slide. After drying, use a glass pen to draw a line on both sides of the coated area (to limit the flow of staining solution). Inside the line, add 3~4 drops of staining solution, after 3~5 minutes, then add an equal amount of distilled water, and shake gently to mix. After 5 minutes, rinse with distilled water and leave to dry before checking with an oil mirror.

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