Tissue samples were rinsed with 1× phosphate-buffered saline (1× PBS) and fixed in 4% paraformaldehyde for 24 h. Samples were then undergone gradual dehydration with sucrose and embedded in 20% gelatin in 1× PBS and stored at – 80 °C. Twenty-micron thick sections were cut using a cryo-microtome (MICROM HM550, Thermo, U.S.A.), and sections were stained with either Giemsa stain (SIGMA, U.S.A.) for tissue integrity or Alcian Blue stain (ScyTek, U.S.A.) for labeling goblet cells. All the prepared slides were stored at 4 °C for later light microscopy examination.
Giemsa stain
Manufactured by ScyTek Laboratories
Sourced in United States
Giemsa stain is a laboratory reagent used for the differential staining of blood smears and other cytological preparations. It is an important tool in the field of hematology and parasitology, providing a reliable method for the identification and differentiation of various blood cells and parasites.
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2 protocols using giemsa stain
1
Tissue Sectioning and Staining
2
Tissue Sectioning and RNA Extraction
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Tissue samples were rinsed with 1X phosphate-buffered saline (1X PBS) and xed in 4% paraformaldehyde for 24 h. Samples were then undergone gradual dehydration with sucrose and embedded in 20% gelatin in 1X PBS and stored at -80℃. Twenty-micron thick sections were cut using a cryo-microtome (MICROM HM550, Thermo, U.S.A.), and sections were stained with either Giemsa stain (SIGMA, U.S.A.) for tissue integrity or Alcian Blue stain (ScyTek, U.S.A.) for labeling mucous cells. All the prepared slides were stored at 4℃ for later light microscopy examination.
Total RNA extraction and cDNA preparation RNA from the skin of P. hypophthalmus was extracted with the TriPure Isolation Reagent (Roche, Mannheim, Germany) following the manufacturer's instructions. The RNA pellet was dissolved in nuclease-free water. Extracted RNA samples were stored at - 80°C. The reverse transcription was then performed using M-MLV Reverse transcriptase (Promega, U.S.A.) to synthesize cDNA following the manufacturer's instructions. The successful construction of cDNA library was determined by 1.5% agarose gels containing the Safeview DNA stain (GeneMark, Taipei, Taiwan).
Total RNA extraction and cDNA preparation RNA from the skin of P. hypophthalmus was extracted with the TriPure Isolation Reagent (Roche, Mannheim, Germany) following the manufacturer's instructions. The RNA pellet was dissolved in nuclease-free water. Extracted RNA samples were stored at - 80°C. The reverse transcription was then performed using M-MLV Reverse transcriptase (Promega, U.S.A.) to synthesize cDNA following the manufacturer's instructions. The successful construction of cDNA library was determined by 1.5% agarose gels containing the Safeview DNA stain (GeneMark, Taipei, Taiwan).
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