Testis integrity was analyzed by fixing testes from WT and NR_038002-KO male mice by immersion in Bouin’s solution overnight. The tissue was then dehydrated, embedded in paraffin, and sectioned (5 μm thick). Testis sections from male WT and NR_038002-KO mice were H&E stained and scanned using an Aperio ScanScope CS2 System (Leica Biosystems). For sperm counts, sperm from the cauda epididymis and vas deferens of male WT and NR_038002-KO mice were collected and counted in a hemocytometer under a light microscope. For analysis of sperm head morphology, mature sperm were isolated from the cauda epididymis. Sperm from male WT (n = 8) and NR_038002-KO (n = 8) mice were spread onto microscope slides and allowed to air dry. About 200 sperm on each slide were analyzed under a light microscope. For fertility tests, adult C57BL/6 WT females (8 weeks old) were mated with sexually mature male WT (n = 10) and NR_038002 KO (n = 8) mice for 14 days and checked daily for vaginal plugs. The fertility rate was calculated from the number of fetuses produced by pregnant females, counted 12 to 16 days after mating.
Aperio scanscope cs2 system
Manufactured by Leica
Sourced in Italy, Germany
The Aperio ScanScope CS2 System is a digital pathology slide scanner designed for high-throughput scanning of microscope slides. The system is capable of capturing high-resolution digital images of tissue samples, providing a comprehensive digital representation of the original specimen.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Anti rabbit alexa488, by Thermo Fisher Scientific (1 mentions)
Anti cd11b alexa647, by BioLegend (1 mentions)
Anti cd34 clone mec14, by BioLegend (1 mentions)
Anti cd3 clone sp7, by Abcam (1 mentions)
Anti cd45r b220 clone ra3 6b2, by BD (1 mentions)
4 protocols using aperio scanscope cs2 system
1
Analysis of Male Fertility in NR_038002-KO Mice
2
Immunohistochemical Analysis of Tissue Macrophages
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Organs were harvested, fixed in zinc formalin, processed, embedded in paraffin, sectioned, and either stained with hematoxylin–eosin (HE) or were further processed for immunohistochemical analysis, as previously described [41 (link)]. Immunohistochemical staining for tissue-resident monocytes/macrophages was conducted with the anti-F4/80 specific antibody (clone A3-1, AbD Serotec, Luxembourg, Luxembourg). All images were acquired using the Aperio Scanscope CS2 system (Leica Biosystems, Milano, Italy), available at the San Raffaele Advanced Light and Electron Microscopy BioImaging Center (ALEMBIC, Milano, Italy). Images were identified as representative areas of interest within the total area of the specimen, then analyzed and exported as ImageScope snapshots.
3
Immunohistochemical and Immunofluorescence Analysis
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At the time of autopsy, different organs for each mouse were sampled and either fixed in zinc–formalin or 4% paraformaldehyde and processed as described in Appendix Supplementary Materials and Methods . Immunohistochemical staining was performed utilizing the following antibodies: anti‐F4/80 (clone A3‐1, AbD Serotec); anti‐RFP (rabbit polyclonal, ab62341 AbCam); anti‐CD34 (clone MEC14.7, Biolegend); anti‐Ki67 (clone SP6, Neomarkers); anti‐CD3 (clone SP7, AbCam); and anti‐CD45R/B220 (clone RA3‐6B2, BD Pharmingen). All images were acquired using the Aperio Scanscope CS2 system (Leica Biosystems). Immunofluorescence staining was performed utilizing the following antibodies: anti‐GFP (rabbit polyclonal, A11122 Invitrogen) + anti‐rabbit Alexa 488 (Invitrogen); anti‐MMR (goat polyclonal, AF2535 R&D Systems) + anti‐goat Alexa 647 (Invitrogen); anti‐F4/80‐PE (clone A3‐1, AbD Serotec); anti‐CD11b‐Alexa 647 (clone M1/70; Biolegend); and Hoechst 33342 (Invitrogen). Confocal images were acquired using a Leica TCS SP2 or SP8 confocal system (Leica Microsystems) that are available at the SRSI Advanced Light and Electron Microscopy BioImaging Center (ALEMBIC).
4
Histological Analysis of Ethanol Effects
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To assess the effect of ethanol administration, we performed histological analysis of mouse testis and liver (caudate lobe) samples. Tissues were fixed and dehydrated using Bouin's solution (for testis), or 3.7% formaldehyde solution (for liver), paraffin embedded, and sectioned at 4 mm. Sections were stained with hematoxylin and eosin (H&E), and whole slides were scanned with an Aperio ScanScope CS2 System (Leica Biosystems, Wetzlar, Germany). The histological sections were analyzed from three mice per group.
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