Paraffin section neuron Golgi method rapid staining kit instruction manual

Paraffin section neuron Golgi method rapid staining kit product manual (Chinese version)

The main purpose

Paraffin section neuron Golgi method rapid staining reagent is a method for analyzing neuron (neuron), pyramidal cell, glial cell (glia cell) in fixed-process paraffin-embedded tissue sections using pro-silver reduction technique. ), the oligodendrocyte and other processes, especially the authoritative and classical technical methods of dendrites morphology. This technology has been proven by the meticulous improvement of the Golgi-Cox method and successful experiments. It is mainly used for the detection of related nerve cells in paraffin sections of brain tissue or peripheral nerve tissue. It is widely used in the study of pathophysiology and anatomy of animal brain. The product is strictly sterile, ready to use, simple in operation, stable in performance and clear in color.

technical background

In 1873, the Italian scientist Camillo Golgi invented the immersion silver staining technique for the black reaction of nerve cells, which opened up the study of the morphology, evolution, monophylogenesis, construction and disease of the nervous system, and established the theory of neurology. The brain nerve tissue was fixed, and after the silver staining, some nerve cells were completely stained, while other nerve cells were not highly stained with highly selective staining.

product content

Fixative A (Reagent A) ml

Fixative B (Reagent B) ml

Dewaxing liquid (Reagent C) ml

Replenishing liquid A (Reagent D) ml

Replenishing liquid B (Reagent E) ml

Replenishing liquid C (Reagent F) ml

Cleaning solution (Reagent G) ml

Staining solution (Reagent H) ml

Coloring solution A (Reagent I) ml

Coloring Solution B (Reagent J) ML

Product manual 1 copy

storage method

Store in a 4 ° C refrigerator to avoid light; reagents are corrosive, pay attention to operational safety; effective guarantee for March

User-supplied

30% sucrose: used for sample dehydration

Non-ionized water: used for working fluid preparation

1.5 ml centrifuge tube: container for working fluid preparation

Sterile scalpel and forceps: used to dissect animal brain tissue

Small glass dyeing tank: container for tissue or slicing

Slicer: for tissue sectioning

Gelatinized slides and coverslips: used for slicing after slicing

Neutral resin: used for slicing

Optical microscope: used for observation and analysis after sectioning

Experimental procedure

Sample fixation

Conventional anesthesia animals and surgery (recommendation: use physiological saline three times per heart, 60 ml each time, remove blood until clear) immediately remove the brain tissue carefully

Cut the brain tissue into 0.5 cm thick tissue blocks with a sterile scalpel

• Immediately clamp the tissue block with sterile forceps
• Xx mL carefully transferred to a fixing solution A (Reagent A) and fixing ml of solution B (Reagent B) in a mixed solution of xx (20 mL / 2 tissue blocks)
• Incubate for 2 weeks at room temperature to avoid light
• Carefully transfer to a user-supplied 30% sucrose solution
• Incubate at 4 ° C for 48 hours to avoid light (Note: should not exceed 1 week )
• Use tissue paper to dry tissue quickly
• Immediately after conventional paraffin treatment, embedding
• Remove the tissue block, perform slow paraffin sectioning, 20 to 100 microns thick, and lay it on gelatinized slides

Dewaxing

• Remove 10 pieces of 20 to 100 micron thick paraffin-embedded tissue sections to be tested
• Into the small staining tank and incubate according to the following table

• Carefully remove the cleaning solution from the slice ( Reagent G )

• Sample dyeing

Prepare a 20 ml plastic bottle, add 10 ml of user-supplied ion-free water, then add xx ml of Reagent I and Reagent J separately , mix and mark as color developing solution. (Use for 10 days) to avoid lighting. Then do the following.

• Carefully add xx μL of staining solution (Reagent H) to cover the entire surface of the sample
• Incubate for 30 minutes at room temperature
• Carefully remove the staining solution from the section (Reagent H)
• Carefully incubate the sections in xx ml of clean solution ( Reagent G ) for 1 minute at room temperature.
• Carefully remove the cleaning solution from the slice ( Reagent G )
• Carefully add xx microliters of color developing solution to cover the entire surface of the sample
• Incubate for 10 to 30 minutes at room temperature until it appears black, terminate immediately, avoid light (Note: can be observed under the microscope during the period)
• Carefully remove the liquid color work on sections
• Carefully incubate the sections in xx ml of clean solution ( Reagent G ) for 1 minute at room temperature.
• Carefully remove the cleaning solution from the slice ( Reagent G )
• (Selection step) for counterstaining operation (CRESYL VIOLET counterstaining kit - YIJI80052 is recommended)
• Transparent processing
• Put a cover slip or cover (neutral resin)
• Immediately under normal light microscopy: neurons, pyramidal cells, glial cells, less synaptic glial cells (oval like beads) and other protrusions, appearing black (if counterstained - the nucleus appears blue, cytoplasm Nissl (material) material appears pink to purple)

Precautions

• This product is 20 operations
• Wear gloves when handling
• Corrosive reagents, note safe operation, in particular using staining solution (Reagent H)
• The gelatinized slides must be used for the spread, otherwise the dye will fall off: the first brush is used; the second is kept wet for 3 hours; the third is wrapped with PARAFIN and then compressed.
• Slices are recommended to be 100 to 200 microns. Too thick and too thin is not conducive to detecting nerve cells.
• It is recommended to use a glass dyeing cylinder
• Keep the sliced ​​surface basically dry each time the reagent solution is changed
• When the reagent solution is slicing the surface, avoid the presence of air bubbles while ensuring that the surface of the slice is covered.
• The entire operation, in the dark state
• Immediately after the dyeing, optical microscopy
• Save the sample after dyeing to avoid illumination
• The company provides a series of tissue cell nervous system components staining reagent products

Quality Standard

This product has been certified to be stable.

This product has been identified and clearly colored

Functional Oligosaccharide

Functional oligosaccharides refer to oligosaccharides that are difficult or impossible to be digested and absorbed by the human body and have special physiological effects on the human body. Its sweetness is generally only 30-50% of that of sucrose, and it has physiological functions such as low-calorie, anti-caries, prevention and treatment of diabetes, and improvement of intestinal colony structure. Due to the special physiological functions of functional oligosaccharides, it becomes a nutrient and health care product. A new generation of food-effect raw materials that integrate diet and therapy. It is a new functional sugar source that replaces sucrose and has a wide range of uses and application prospects. Common functional oligosaccharides include: xylo-oligosaccharides, fructooligosaccharides, galacto-oligosaccharides, isomaltose, raffinose and so on.

XOS, GOS, FOS, IMO, Raffinose, oligosaccharide

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