Cytospins were performed following standard procedures. Briefly, cell suspensions from dissociated granulomas were spun for 3 minutes at 800 rpm in a Shandon Cytospin3 apparatus. Slides were dried overnight at room temperature, and subsequently fixed in absolute methanol for 1 minute, stained with Wright-Giemsa (Sigma, WG16) for 1 minute, rinsed in ddH2O, then coverslipped with DPX mounting media (Sigma, 06522). Slides were imaged on a Zeiss AxioObserver Z.1 with an 40×/0.75 NA (Zeiss, EC Plan-Neofluar) and an ICc5 color camera (Zeiss) using Zen Blue 2012 edition (Zeiss).
Icc5 color camera
Manufactured by Zeiss
Sourced in Germany
The ICc5 color camera is a laboratory-grade camera developed by Zeiss. It is designed to capture high-quality color images for various scientific and industrial applications. The camera features a CMOS sensor and offers a range of resolution options to meet the needs of different imaging requirements.
Automatically generated - may contain errors
3 protocols using icc5 color camera
1
Cytospin Protocol for Granuloma Analysis
2
Pollen Grain Vitality Analysis
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Anthers were fixed with FAA fixative and stained with DAPI, periodic acid-Schiff (PAS), and fluorescein diacetate (FDA) following previously described methods [29 (link),59 (link)]. The FDA assay was performed to assess the vitality of fresh pollen grains. 3,3′-diaminobenzidine (DAB) staining with the diaminobenzidine method was conducted using 1 mL of 10 mM Tris-HCL, DAB, and 0.03% CoCL2 mixed, to which 10 mL of 30% H2O2 was added, after which the solution was mixed again. Anthers were placed in DAB staining solution in dark incubation for 3 h and then placed in deionized water for cleaning. Mature pollen was stained with I2-KI and peroxidase (following the aniline method). Images of the microspores were acquired using ICc5 color camera (ZEISS, Oberkochen, Germany) mounted onto a biological fluorescence microscope (ZEISS Imager M2, Oberkochen, Germany). Images of the anther and the seed status after wheat pollination were acquired using a stereoscopic microscope (OLYMPUS SZX16, Tokyo, Japan).
3
Cytospin Protocol for Granuloma Analysis
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