Cryostat sections were obtained from bowel specimens perfused with O.C.T. compound and snap frozen. After warming the slide to 27 °C, the sections were fixed for 10 min (2% paraformaldehyde and PBS at pH 7.43). The slides were washed with buffer (PBS, 5% sheep serum, 0.1% azide, 1 mM MgCl2, 1 mM CaCl2) and blocked with 20% sheep serum in PBS. The slides were treated with the biotinylated lectin followed by avidin-fluorescein (Southern Biotech, Birmingham, AL, USA) or avidin-fluorescein alone control. The slides were incubated for 1 h at 27 °C, washed 3 times, and mounted with DAPI-containing medium (Vector Labs, Burlingame, CA, USA). The tissue sections were imaged with a Nikon Eclipse TE2000 inverted epifluorescence microscope (Tokyo, Japan).
Eclipse te2000 inverted epifluorescence microscope
Manufactured by Nikon
Sourced in Japan
The ECLIPSE TE2000 is an inverted epifluorescence microscope manufactured by Nikon. It is designed for various applications that require fluorescence imaging. The microscope features a stable, ergonomic design and offers high-quality optics for bright, clear images.
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Lab products found in correlation
3 protocols using eclipse te2000 inverted epifluorescence microscope
1
Lectin-based Fluorescent Staining of Bowel
2
Visualizing Primordial Germ Cells in Mice
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Embryos were harvested at E9.5, E10.5 and E11.5 following timed matings between Ercc1 heterozygous mice carrying the GOF18-GFP reporter, as described previously. Embryos were fixed in 10% neutral-buffered formalin overnight at 4 °C and washed three times in PBS for five minutes each. In the case of E10.5 embryos the right body wall was dissected to reveal the genital ridge and placed on their left side and embedded in 1% w/v low melting point agarose. In the case of E11.5 embryos the front abdominal wall and visceral organs were dissected away to reveal the genital ridge47 (link) and embryos were placed ventral side down and embedded in 1% w/v low melting point agarose to facilitate visualization of GOF18-GFP+ PGCs. Samples were incubated at 4 °C for 30 minutes, until the agarose had fully set. Whole-mount fluorescence images were acquired using an ECLIPSE TE2000 inverted epifluorescence microscope (NIKON). In the case of E10.5 embryos somites were used as physiological landmarks to assess the distribution of PGCs along the hindgut.
3
Visualizing Primordial Germ Cells in Mice
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