Immunofluorescence staining was adopted to assess the expression level of chondrocyte markers type I collagen (Col1), type II collagen (Col2), and type X collagen (Col10). After differentiation of the cells at days 7, 14, and 21, the cells were fixed in 4% paraformaldehyde, followed by permeabilization with 0.5% Triton X-100 in phosphate-buffered saline (PBS). After being washed, the cells were blocked with 1% bovine serum albumin (BSA) in 0.3 M of glycine and incubated with primary antibody against Col1 (AB34710 at 1:500 dilution, Abcam, Cambridge, United Kingdom), Col2 (AB34712 at 1:250 dilution, Abcam, United Kingdom), and Col10 (AB49945 at 1:1,000 dilution, Abcam, United Kingdom) overnight at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG and an Alexa Fluor® 594-conjugated goat anti-mouse IgG were used as the secondary antibody. The cellular nuclei were counter-stained with DAPI (Sigma, USA). The immunofluorescence was observed with a Nikon-Ti inverted fluorescence microscope (Nikon, Tokyo, Japan).
Ti inverted fluorescence microscope
Manufactured by Nikon
Sourced in Japan
The Nikon Ti inverted fluorescence microscope is a high-performance laboratory instrument designed for advanced fluorescence imaging applications. It features a motorized inverted configuration, which allows for the observation of samples from the underside. The microscope is equipped with a variety of fluorescence illumination options, including LED and mercury-based light sources, enabling researchers to capture detailed images of fluorescently labeled specimens.
Automatically generated - may contain errors
Lab products found in correlation
80i upright microscope, by Nikon (2 mentions)
Vimentin, by Cell Signaling Technology (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (2 mentions)
Ab49945, by Abcam (1 mentions)
Ab34712, by Abcam (1 mentions)
Ab34710, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Primary antibodies against β catenin, by Abcam (1 mentions)
Ab45939, by Abcam (1 mentions)
Ab76055, by Abcam (1 mentions)
Ab27568, by Abcam (1 mentions)
Ab205270, by Abcam (1 mentions)
E cadherin, by Abcam (1 mentions)
Nis element, by Nikon (1 mentions)
Readyprobes cell viability imaging kit blue green, by Thermo Fisher Scientific (1 mentions)
Snai2, by Cell Signaling Technology (1 mentions)
Alexa 594 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions)
Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
26 protocols using ti inverted fluorescence microscope
1
Chondrocyte Marker Expression Analysis
2
Telomere Loss Detection in GSCs
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FISH was used to detect the telomere loss. GSCs were collected and treated with 10 μg/mL colcemid for 20 h, incubated with 75 mM KCl for half an hour at 37°C, prefixed for 5 min and then fixed for 1 h with a mixture of methanol and glacial acetic acid (3:1). The cells were dropped onto water-coated slides to spread the metaphase chromosomes. Then, the slides were stained with the PNA probe following the manufacturer's protocol. Images were captured by a Nikon Ti inverted fluorescence microscope. The images were analyzed using NIS Elements Advanced Research software.
3
Telomere Damage Analysis in GSCs
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Telomere damage was analyzed with IF-FISH. GSCs were plated onto poly-L-lysine-coated glass coverslips in DMEM/F12 for 1 h and then fixed with 4% paraformaldehyde for 20 min. The cells were permeabilized with 0.2% Triton X-100 (Sigma, USA) and dehydrated through graded alcohol. The coverslips were hybridized with the PNA probe, 53 bp1 antibody and secondary antibody following the manufacturer's protocol. Images were acquired on a Nikon Ti inverted fluorescence microscope. The images were deconvoluted and analyzed using NIS Elements Advance research software. The foci counts were exported to Excel for analysis.
4
Immunofluorescence Staining of β-Catenin
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After the 1 h incubation of CC cells with blocking buffer (5% normal goat serum, 3% bovine serum albumin and 0.1% Triton-X 100 in PBS), the primary antibodies against β-catenin (Abcam, Cambridge, U.K.) were added and incubated with the CC cells overnight. Following rinsing for three times with PBST, cells were then incubated with secondary antibodies for 1 h under room temperature. DAPI (Burlingame, CA) was then used to stain the cell nuclei. Pictures were taken applying the Nikon Ti inverted fluorescence microscope.
5
Imaging Techniques for Stained Biological Samples
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In O-Dianisidine staining assays, stained embryos were imaged and photographed using a Leica M80 Microscope with a Nikon D200 digital camera using an adjustable flash system. Erythrocytes with May-Grünwald/Giemsa staining were imaged by Nikon 80i Upright Microscope in Nikon Image Center at Harvard Medical School. Fluorescence and/or DAPI stained images were taken with a Nikon Ti Inverted Fluorescence Microscope with Perfect Focus System in Nikon Image Center at Harvard Medical School. Images of zebrafish embryos were taken by a Leica M80 Microscope with a Nikon D200 digital camera.
6
Immunofluorescence Analysis of Ovarian Cancer Markers
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Ovarian cancer cells were incubated with blocking buffer (5% normal goat serum, 3% bovine serum albumin, and 0.1% Triton-X 100 in PBS) for 1 h. Primary antibodies to YAP1 (ab205270; 1:200; Abcam), E-cadherin (ab76055; 1:150; Abcam), vimentin (ab45939; 1:100; Abcam), Slug (ab27568; 1:150; Abcam), and PCNA (ab29; 1:100; Abcam) were incubated with the cells overnight. After three rinsing for 5 min with PBST, Alexa Fluor 568 (red, goat anti-rabbit), 488 (green, goat anti-rabbit) and 488 (green, goat anti-mouse) antibodies were applied (1200 dilution, Invitrogen, Thermo Fisher, USA) for 1 h at room temperature. Cell nuclei were counterstained with DAPI (Burlingame, CA). Images were taken using a Nikon Ti inverted fluorescence microscope.
7
Imaging Techniques for Stained Biological Samples
8
Confocal and TIRF Imaging Protocols
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Confocal imaging of brain slices was done using LSM710 Duo imaging system (Zeiss, Thornwood, NY,USA) with either a × 20 non-immersion or × 63/1.4 Plan-Apochromat oil immersion objectives, as described in Supplementary Methods . Total internal reflection fluorescence (TIRF) imaging of the bottom plane of MEFs was done using a Nikon Ti inverted fluorescence microscope with a × 60/1.49 Apo-TIRF oil objective (Nikon) and a filter set for GFP illumination.
9
Imaging Macrophage Response to S. aureus
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Macrophages were challenged with S. aureus Newman‐GFP strain at an MOI of 5 for 6 h or mock infected, and extracellular bacteria were killed with lysostaphin. Macrophages were then incubated in fresh media without sodium hydrogen carbonate with or without lysostaphin for up to 72 h. Imaging was started at 52 h post infection, imaging every 10 min until 72 h. Images were taken using the ×30 DIC/GFP laser on the Nikon Ti inverted fluorescence microscope with a 20 × 0.75 NA lambda objective lens. Images were captured with Neo camera (Andor) using NIS Elements (Nikon). The microscope was enclosed in temperature‐controlled and humidity‐controlled cabinet (OKO Labs) maintained at 37 °C. Images were taken from four fields of view from three random wells and analysed using the NIS Elements Viewer software version 4.20 (Nikon).
10
Immunofluorescent Analysis of Ovarian Tumor Markers
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