Frozen section neuron Golgi staining kit instructions

Frozen section neuron Golgi staining kit product manual (Chinese version)

The main purpose

YIJI Frozen Slice Neuron Golgi staining reagent is a method for analyzing neuron (neuron), pyramidal cell, glia cell, and frozen tissue sections in fixed treatment using pro-silver reduction technique. An authoritative and classical technique for the morphological study of oligodendrocytes and other processes, especially dendrites. The technology has been proven through careful improvement of traditional methods and successful experiments. It is mainly used for the detection of related nerve cells in frozen brain tissue or peripheral nerve tissue sections. 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

YIJI Fixative (Reagent A) ML

YIJI oxidizing solution (Reagent B) ml

YIJI Nutrient Solution (Reagent C) ml

YIJI Reinforced Liquid (Reagent D) ML

YIJI Dehydrated Liquid (Reagent E) ML

YIJI cleaning solution (Reagent F) ml

YIJI staining solution (Reagent G) ml

Product manual 1 copy

storage method

Store in a 4°C refrigerator to avoid light; valid for March

User-supplied

1.5 ml centrifuge tube: container for working fluid preparation

Sterile scalpel and forceps: used to dissect animal brain tissue

Small glass dyeing tank or 60mm glass petri dish: container for tissue slicing

Collodion: used to protect tissue samples after fixation

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 to infuse the heart three times, 60 ml each time, remove blood until clarification)
• Appropriate amounts YIJI fixing solution (Reagent A) drenching
• Immediately remove the brain tissue carefully
• Carefully put into xx ml of YIJI Fixative ( Reagent A )
• Incubate for 1 hour at room temperature
• Cut the brain tissue into 0.5 cm thick tissue blocks with a sterile scalpel
• Immediately clamp the tissue block with sterile forceps
• Carefully transfer to xx ml of YIJI oxidant ( Reagent B ) (20 ml/2 block)
• Incubate for 2 weeks at room temperature to avoid light
• Carefully transfer to xx ml of YIJI Nutrient Solution ( Reagent C )
• Incubate for 48 hours at room temperature to avoid light
• Carefully transfer to xx ml of YIJI fortification ( Reagent D )
• Incubate for 48 hours at room temperature to avoid light (Note: can be incubated for 1 week )
• Use tissue paper to dry tissue quickly
• Embedded with 30% sucrose, embedded in OCT or embedded with user-supplied collodion (8% collar)
• Put it in the refrigerator at -20 °C
• The tissue block was removed and slowly frozen at -20 ° C for 20 to 100 μm thick and plated on gelatinized slides.
• Store in a -20 ° C refrigerator

• Sample dyeing

• Take out 20 to 100 micron thick frozen tissue sections to be tested
• Carefully add xx μl of YIJI Dehydrate ( Reagent E ) on the slice to cover the entire sample surface.
• Incubate for 2 minutes at room temperature
• Carefully remove the YIJI dewatering solution ( Reagent E ) on the slice
• Repeat experiment steps 2 to 4
• At room temperature, carefully place the sections in xx ml of YIJI Cleaner ( Reagent F ) for 2 minutes.
• Carefully remove the YIJI cleaning solution on the slice ( Reagent F )
• Carefully add xx microliters of YIJI staining solution (Reagent G) to cover the entire surface of the sample.
• Incubate for 48 hours at room temperature until it appears black, immediately terminate, avoiding light (Note: can be observed under the microscope during the period; avoid drying)
• Carefully remove the YIJI stain on the slice (Reagent G)
• At room temperature, carefully place the sections in xx ml of YIJI Cleaner ( Reagent F ) for 2 minutes.
• Carefully remove the YIJI cleaning solution on the slice ( Reagent F )
• (Selection step) for counterstaining operation (recommended using YIJI cresyl violet (CRESYL VIOLET) counterstaining kit - YIJI80052)
• 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
• 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.
• Some tissues are easily broken by frozen slices. It is recommended to adjust the slice temperature first; the second slice thickness is above 150 microns; if it cannot be solved, it is recommended to use paraffin sections, etc.
• 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

Automatic Nucleic Acid Extractor

Nucleic acid extraction instrument is an instrument that automatically completes the nucleic acid extraction of samples by applying the supporting nucleic acid extraction reagents. Widely used in disease control centers, clinical disease diagnosis, blood transfusion safety, forensic identification, environmental microbial testing, food safety testing, animal husbandry and molecular biology research and other fields.
Using magnetic beads as the carrier, using the principle that magnetic beads adsorb nucleic acid under high salt and low pH value, and separate from nucleic acid under low salt and high pH value, and then move the magnetic beads or transfer the liquid to realize the entire extraction and purification process of nucleic acid. Due to the uniqueness of its principle, it can be designed into a variety of throughputs, which can be extracted from a single tube or 8-96 samples, and its operation is simple and fast. It only takes 30-45 minutes to extract 96 samples, which greatly improves the The experimental efficiency is low and the cost is low, so it can be used in different laboratories, and it is the mainstream instrument on the market.

Features of Magnetic Bead Nucleic Acid Extractor
1. Enables automated, high-throughput operations.
2. Simple and fast operation.
3. Safety and environmental protection.
4. High purity and high yield.
5. No pollution and stable results.
6. Low cost and easy to be widely used.
7. Different types of samples can be processed simultaneously.

Nucleic Acid Extraction System,Automatic Nucleic Acid Extractor,Nucleic Acid Extraction Machine,Nucleic Acid Extraction Instrument

Jilin Sinoscience Technology Co. LTD , https://www.contoryinstruments.com