Digital sight ds fi camera

Day 5 expanded, hatching and fully hatched blastocysts were selected for differential nuclear staining at 121 h post-hCG to assess the number of inner cell mass (ICM) and trophectoderm (TE) cells (Hardy et al., 1989 (link)). Pronase was used to remove the zona, after which 10 mM trinitrobenzenesulphonic acid, 0.1 mg/ml anti-dinitrophenol and guinea pig complement serum containing 0.1 mg/ml propidium iodide were utilized to label TE cells, while the ICM cells remained unlabelled. All cells were counterstained in 0.1 mg/ml bisbenzimide (Hoechst 33258). Blastocysts were mounted on glass microscope slides in 100% (v/v) glycerol and imaged on a Nikon Eclipse Ts100 inverted fluorescent microscope equipped with a Nikon digital sight DS-Fi camera. FIJI image analysis software (ImageJ 1.52a, https://imagej.net/software/fiji/) was used to count cell numbers.

Wet tissues were collected in embedding cassettes and formalin-fixed in 4% neutral buffered formaldehyde (Engelbrecht, Edermünde, Germany) at room temperature for about 24 h, followed by automatic dehydration and embedding in IHC-grade paraffin using a Leica TP 1020 (Leica Biosystems, Nussloch, Germany) and Leica Histo Core Arcadia H/C (Leica Biosystems). FFPE tissue blocks were stored at room temperature. Sections from paraffin blocks with non-specified tissues were cut into slices of approximately 2–3 µm thickness and routinely stained with HE at TPL Path Labs (Freiburg, Germany).
Histopathological examinations were then performed blind using a Zeiss Axioscope microscope (Carl Zeiss, Jena, Germany) at a magnification of up to 400×, by one of the authors (T.L.). Digital microphotographs were taken using a Nikon Digital Sight DS-Fi camera (Nikon Instruments Europe B.V., Amsterdam, Netherlands). Whole slide imaging using an Axioscan Z1 (Carl Zeiss) were also performed from all case, for the purpose of comprehensive iconography.

Wet tissues were collected in embedding cassettes and formalin-fixed in 4% neutral buffered formaldehyde (Engelbrecht, Edermünde, Germany) at room temperature for about 24 h, followed by automatic dehydration and embedding in IHC-grade paraffin using a Leica TP 1020 (Leica Biosystems, Nussloch, Germany) and Leica Histo Core Arcadia H/C (Leica Biosystems). FFPE tissue blocks were stored at room temperature. Sections of the paraffin blocks were cut into slices of approximately 2–3 µm thickness and stained with H&E at TPL Path Labs (Freiburg, Germany).
Histopathological examinations were then performed blind using a Zeiss Axioscope microscope (Carl Zeiss, Jena, Germany) at a magnification of up to 400×, by one of the authors (TL). Digital microphotographs were taken using a Nikon Digital Sight DS-Fi camera (Nikon Instruments Europe B.V., Amsterdam, Netherlands). Whole slide imaging using an Axioscan Z1 (Carl Zeiss) were also performed from all case, for the purpose of comprehensive iconography.