Eight array slides with 192 cores were stained with immunofluorescence. Using multiplex imaging, slides were processed using the staining protocol above until after protein block, at which point arrays were wet mount with 50/50 glycerin/PBS and 1:5000 DAPI for imaging on a Leica Versa 200 to create digital H&E with a whole slide fluorescent scanning platform. Slides were decoverslipped in PBS and primary antibodies for TFF3, Trop2, CD44v9, and PCNA were incubated overnight at 4°C following staining protocol as described above. Species-specific secondary antibodies were incubated for 1 hour at room temperature for Cy2, Cy3, Cy5, and Alexa 750. Slides were wet mounted with glycerin/PBS/Hoechst solution and imaged on the Leica Versa 200 imager for immunofluorescence. Three cores were lost during the staining process. Staining index in cancer regions for Trop2, CD44v9 and TFF3 images was stratified as follows: 0 = no staining, 1 = staining in up to 25% of cells, 2 = in 25–50% of cells, 3 = staining in greater than 50% of cells.
Versa 200
Manufactured by Leica
Sourced in Germany
The Leica Versa 200 is a high-performance lab equipment designed for versatile imaging applications. It features a cutting-edge optical system and advanced imaging capabilities to deliver precise and detailed visual information.
Automatically generated - may contain errors
Lab products found in correlation
Z 1 light sheet microscope, by Zeiss (1 mentions)
Bz x810 widefield microscope, by Keyence (1 mentions)
Deadend fluorometric tunel system, by Promega (1 mentions)
Scn400, by Leica (1 mentions)
Refine detection kit, by Leica (1 mentions)
Bond rx, by Leica (1 mentions)
Clone d2p2f, by Cell Signaling Technology (1 mentions)
P tbk1, by Cell Signaling Technology (1 mentions)
Sting, by Cell Signaling Technology (1 mentions)
9 protocols using versa 200
1
Multiplex Immunofluorescence Staining Protocol
2
Multimodal Microscopy Techniques for Embryonic Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H&E-stained embryos were imaged with Leica Aperio VERSA 200 (40× objective). ISH and immunofluorescence-stained sections were imaged with Olympus FV3000S laser-scanning confocal microscope (40× [NA 1.25] and 60× [NA 1.25] objectives). Whole-mount–stained embryos were imaged by Zeiss Z-1 Lightsheet Microscope (5× [NA 0.16] objective). Immunolabeled embryo sections and cultured cells were imaged with Keyence BZ-X810 Widefield Microscope (10× [NA 0.45], 20× [NA 0.75], and 40× [NA 0.95] objectives). Filipin-labeled cultured cells were imaged with Keyence BZ-X810 Widefield Microscope (60× [NA 1.4] oil objective). The proliferation of NSCs was imaged with Incucyte S3 Live Cell Imaging system (10× [NA 0.3] objective) and analyzed with S3 2020A GUI software. The Incucyte system uses a 12-bit CMOS camera (scientific grade CMOS camera) and contrast-based autofocus to acquire images. Nile Red stained cells were imaged with Zeiss Elyra 7 for Lattice SIM Microscopy with 405, 488, and 561 nm lasers, PlanApo 63× oil immersion objective (NA 1.40), and acquired using a dual camera: 2 pco.edge 4.2 camera system with lattice sim leap mode. Subsequently, Nile Red images were processed using Zeiss’s automated sim leap mode pipeline. See Table S4 for microscope details.
3
Apoptosis Quantification in Granulosa Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis induction was analyzed using the DeadEnd Fluorometric TUNEL System (Promega, USA). According to the manufacturer's instructions, each section was incubated with TUNEL reaction mixture at 37 °C for 1 h. The green fluorescence of the apoptotic GCs was examined using Versa 200 (Leica, Germany). The number of TUNEL-positive GCs was calculated using Image-Pro Plus.
4
Immunostaining of Dysplastic Adenomas
5
Immunohistochemical Analysis of Tumor Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumor tissues were isolated, fixed in 10% formalin, embedded in paraffin, and cut into 4 μm paraffin-embedded sections for IHC and HE assay. For IHC, sections were subjected to antigen retrieval in 10 mmol/L sodium citrate buffer (pH 6.0) in a microwave oven. Sections were then incubated with the primary antibody at 4 ℃ overnight and with a biotinylated secondary antibody for 30 min, followed by the avidin–biotin–peroxidase complex for another 10 min. Staining was visualized using 3,3′-diaminobenzidine solution (Maxim, Fuzhou, China). For HE, sections were stained with hematoxylin-eosin, and images were obtained with Leica Aperio Versa 200. Immunohistochemical staining was blindly scored by two pathologists, and the average score was taken as the final result. The percentage of positive tumor cells was determined in at least five areas and scored as the following: 0, ≤5%; 1, 5–25%; 2, 25–50%; 3, 50–75%; 4, ≥75%. The intensity of staining in positive cells was scored as 1 (weak), 2 (moderate), or 3 (intense). The total score was calculated by multiplying the scores for percentage and the intensity of staining.
6
Fluorescence Imaging of Mouse Gastric Organoids
7
Lung Histopathology in Mouse Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For histopathology, lung tissues from mice were fixed in 4% neutral-buffered formalin for 48 h, embedded in paraffin, and stained with hematoxylin and eosin (H&E). Images were captured using a LEICA Versa 200 and were processed using software K-Viewer 1.5.5.8.
8
Histopathological Analysis of SARS-CoV-2 Lung Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The lungs were fixed in 4% (v/v) paraformaldehyde solution for 72 h, and the paraffin sections (3–4 μm) were prepared routinely. Hematoxylin and eosin stain was used to identify histopathological changes in the lungs. The histopathology of the lung tissue was observed by light microscopy. For immunohistochemistry, SARS-CoV-2 N protein was detected using monoclonal antibody clone 019 (Sino Biological, China). Images were captured using a LEICA Versa 200 and were processed using software HALO v3.1.1076.379.
9
Multiplex Immunofluorescent Staining Automation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescent multiplex staining was performed on the Leica Bond RX automated staining platform with a Leica Biosystems Refine Detection Kit. Antibodies were used as follows: pTBK1 (Cell Signaling Technologies; clone D52C2, catalog no. 5483) was run at 1:50 dilution with EDTA antigen retrieval; STING (Cell Signaling Technologies; clone D2P2F, catalog no. 13647S) was run at 1:50 dilution with citrate antigen retrieval; CD31 (Cell Signaling Technology; clone D8V9E, catalog no. 77699) was run at 1:100 dilution with citrate antigen retrieval; IBA1 (Wako; polyclonal, catalog no. 019-19741) was run at 1:500 dilution with citrate antigen retrieval. Imaging was performed on a Leica Versa 200 automated fluorescent/brightfield scanner at 20× magnification. Alexafluors 488, 555, 594, and 647 were used for each antibody (respectively).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!