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General blood tests
Basic clinical testing techniques include a detailed summary of blood smear preparation, blood smear staining, cow-type counting boards, and microscopes. Friends, let’s learn quickly~
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General blood tests
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General blood tests
Blood smear preparation
Slide cleaning
New slides: Soak in dilute acid (remove alkali), remove grease
Old glass slides: boil and wash with detergent
Production method
Manual pushing method
Thin blood film smear method: observing blood cells
Thick blood film smear method: observing malaria parasites
automatic smear method
Purpose of film production: blood cells are dispersed for easy observation under the microscope
Blood drop: millet grain size, 5μL±; Angle: 30°-45°; Speed: forward at a constant speed, ending naturally
The faster it is, the thicker it is; the slower it is, the thinner it is.
The role of methanol
Coagulates proteins in cells and acts as a fixation
Increase cell membrane permeability and dissolve dyes
Precautions
Pick marking and selection
Mix the specimen
blood drop size
Angle control
speed control
Criteria for a good blood smear
Appropriate thickness and even distribution
The head, body and tail are distinct, and the tail is feather-shaped without any scratches.
The blood film is at least 25mm long and occupies 1/3-2/3 of the slide.
The edges are neat, leaving gaps on both sides, about 5mm
Most of the white blood cells are intact and evenly distributed. Except for some lymphoproliferative diseases, the amount of WBC damage in the microscopic examination area is <2%
No human pollution
Quality control of blood smear
Blood drop size, film pushing angle, film pushing speed
Causes and Solutions of Poor Blood Smear
blood smear staining
Dyeing method
Wright's stain
Most commonly used, it has good staining effect on cytoplasm.
St. Regis dyeing
Good coloring of cytoplasm and nuclei with sharp contrast
Ginn staining
Good coloring of cell nuclei and parasites
Dye ingredients
Wright's dye: Yin Hong (acidic), Meilan (alkaline)
Gibbs dye: eosin (acidic), azure (alkaline)
Dyeing principle
Chemical affinity, physical adsorption
Purpose of staining: blood cells have different colors for easy identification
Dyeing quality control
Before dyeing
Blood smear: Dye thoroughly before staining, otherwise it will easily fall off during the staining process;
Dye liquor quality
The newly prepared dye must be mature. The more Azure B, the better the dyeing effect.
Wright's dye: the maturity index RA=1.3±0.1 is appropriate
The dye solution is stored in brown bottles to prevent methanol from evaporating or oxidizing into formic acid.
Dyeing
Time and concentration, dyeing process, pH (buffer pH 64~6.8), washing dye solution, destaining and counterstaining
After dyeing
Effect evaluation, reasons for poor dyeing and corrective measures (dye deposition: crayon line drawing to ensure sufficient dye, time, flushing
Bovine counting board
structure
Use (counting principle)
Make it follow a certain direction (such as reverse "S" shape)---prevent missing numbers and repeated numbers
Count two adjacent sides without counting two adjacent sides (for example: count up but not down, count left but not right)
Concentration formula: number of red blood cells/L=N×5×10×201×106
Recharge the pool
equipment
Micropipet + holed tip
principle
Capillary siphon phenomenon (surface tension)
Main points
Maximize surface tension---the counting plates and coverslips are clean and dry
Just fill the counting cell, too little is unevenly distributed, too much overflows---liquid volume 7~8μL
Avoid uneven distribution caused by multiple charging---formation of droplets, charge once
After filling the tank, the cover is fixed and does not move.
microscope
structure
computer system
Lens holder, mirror column, mirror arm, focusing system, lens barrel, nosepiece, stage, press clamp
Optical system
Light source, condenser, shutter, objective lens, eyepiece
use
Focus adjustment
Coarse/fine focus spiral
Light adjustment
Concentrator, shutter, light source
Use of oil lens
Use low/high magnification to select the appropriate location on the smear
Raise the condenser to the highest level and adjust to appropriate brightness
maintain
Silk cloth for wiping mechanical parts and lens paper for optical parts
Wipe the oil lens promptly after use