Representative raw immunofluorescence microscopy images of immune cell markers in the nonpregnant sheep endometrium on days 12, 16, and 19 after estrus

Description

This dataset contains representative immunofluorescence microscopy images of five immune cell-associated markers in endometrial tissues from nonpregnant Kazakh ewes (Ovis aries) on days 12, 16, and 19 after estrus. The markers examined were F4/80, Ly-6G, CD19, CD56, and CD25. For F4/80, Ly-6G, CD19, and CD56, one representative large-field microscopy image is provided at each time point. For CD25, one image is provided at 12D and 16D, and two images are provided at 19D. The dataset therefore contains 19 original ND2 microscopy files and 19 corresponding TIFF images. The images include tissue areas corresponding to the caruncular (C), intercaruncular–caruncular junction (I-C), and intercaruncular (I) regions of the ovine endometrium. Images were acquired using a Nikon A1 HD25 laser-scanning confocal microscope with a 20× objective. ND2 files can be opened using Nikon NIS-Elements or Fiji/ImageJ with the Bio-Formats plugin, whereas TIFF images can be opened using standard image-viewing or image-analysis software. The dataset provides representative nonpregnant reference images for examining the spatial distribution of immune cell-associated markers in the ovine endometrium.

Steps to reproduce

Endometrial tissues were fixed in 4% paraformaldehyde at room temperature for 12 h, processed using standard paraffin-embedding procedures, and sectioned at a thickness of 6 μm. Paraffin sections were deparaffinized in xylene and rehydrated through a graded ethanol series. Antigen retrieval was performed in EDTA antigen retrieval solution (pH 9.0) at 96–97 °C for 16-17 min. After cooling and washing, sections were blocked with 10% goat serum at 37 °C for 60 min and incubated overnight at 4 °C with primary antibodies against F4/80, Ly-6G, CD19, CD56, or CD25 at a dilution of 1:400. For conventional immunofluorescence detection, sections were incubated with a FITC-conjugated goat anti-rabbit IgG secondary antibody at 37 °C for 45 min in the dark. HRP–tyramide signal amplification was used for CD19 and CD25 detection. Nuclei were counterstained with DAPI, and tissue autofluorescence was reduced using 0.3% Sudan Black B. Negative-control sections were processed using PBS instead of the primary antibody. Imag

Institutions

• Xinjiang University

  Xinjiang, Ürümqi

Categories

Funders

• National Key Research and Development Program of China

  Grant ID: 2021YFD12009001
• Key Research and Development Project of Xinjiang Uygur Autonomous Region

  Grant ID: 2023B02039-1
• Project of Tianshan Scholars of Xinjiang

  Grant ID: 2023SNGGGCC021
• Tianshan Cultivation Program of Xinjiang Uygur Autonomous Region

  Grant ID: 2022TSYCLJ0013

Licence