Fluorescence microscopy was performed as previously described 18 (link). Cells were treated with 4% paraformaldehyde for 30 min and then washed twice with PBS. The cells were then treated with 0.1% Triton X-100 for 2 min. After washing twice with PBS, the cells were treated with 1% BSA. They were then incubated with primary antibody. The antibody for detecting ATF4 expression was purchased from Santa Cruz (sc-22800, USA, 1:500). The secondary antibodies (1:200) were conjugated to FITC. DAPI was used for nuclei counterstaining. Immunofluorescent staining of the cells was performed using an Olympus IX51 fluorescent microscope (Olympus, Tokyo, Japan). Images were captured using a CoolPIX 5400 camera (Nikon, Japan) fixed in a fluorescent microscope.
Coolpix 5400 camera
Manufactured by Nikon
Sourced in Japan
The CoolPIX 5400 is a digital camera with a 5.0-megapixel image sensor. It has a 4x optical zoom lens and a 1.8-inch LCD display. The camera supports various image formats, including JPEG and RAW.
Automatically generated - may contain errors
Lab products found in correlation
Ix51 fluorescent microscope, by Olympus (3 mentions)
Ab1416, by Abcam (1 mentions)
Alexa 488 dye, by Thermo Fisher Scientific (1 mentions)
Ix51 fluorescence microscope, by Olympus (1 mentions)
Ab63989, by Abcam (1 mentions)
Sc 81189, by Santa Cruz Biotechnology (1 mentions)
Biorevo bz 9000 fluorescence microscope, by Keyence (1 mentions)
Kl 2500 lcd, by Schott (1 mentions)
Phase contrast light microscope, by Leica (1 mentions)
Live dead viability cytotoxicity kit, by Thermo Fisher Scientific (1 mentions)
6 protocols using coolpix 5400 camera
1
Immunofluorescence Imaging of ATF4 Expression
2
Immunofluorescent Detection of E-cadherin
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Cells were fixed with 4% paraformaldehyde, followed by blocking with 1% BSA. They were then incubated with mouse anti-human monoclonal antibody E-cadherin (ab1416, abcam, HK; dilution 1:100) overnight at 4°C. The primary antibodies were followed by incubation with secondary antibodies conjugated to rhodamine. The nuclei were counterstained with DAPI. The cells were examined with an Olympus IX51 fluorescent microscope (Olympus, Tokyo, Japan), and images were captured with a CoolPIX 5400 camera (Nikon, Japan).
3
Immunofluorescence Staining of Cell Markers
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The MCF‐7 and MDA‐MB‐231 cells were added to a four‐chamber slide glass and incubated overnight. The cells were then washed with phosphate‐buffered saline (PBS) and fixed with ice‐chilled methanol for 30 minutes and permeabilized with 0.2% Triton X‐100 in PBS for 30 minutes. After washing twice with PBS, the permeabilized cells were placed in 5% normal horse serum in PBS for 30 minutes (to block the nonspecific adsorption of antibodies) before incubating with anti‐CLDN1 (1:50), anti‐CLDN4 (1:50), anti‐BHLHE41 (1:50) and anti‐Myc (1:50) antibody at 4°C overnight. The cells were then incubated for 1 hour with Alexa 488 dye or Alexa 488 dye (Molecular Probes, Inc) conjugated to goat anti‐rabbit IgG antibody, while nuclear staining was performed using Hoechst 33258. The cells were observed using an Olympus IX51 fluorescent microscope (Olympus), and the images were captured with a COOLPIX 5400 camera (Nikon).
4
Immunofluorescent Detection of NDRG1 and ATF3
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Cells were fixed with 4% paraformaldehyde and then blocked with 1% BSA, followed by incubation with rabbit anti-human NDRG1 polyclonal antibody (ab63989, Abcam, HK; dilution 1:100) and mouse anti-human ATF3 monoclonal antibody (sc-81189, Santa Cruz, USA; dilution 1:100) overnight at 4°C. The cells were then incubated with secondary antibodies conjugated to rhodamine/fluorescein isothiocyanate (FITC). The nuclei were counterstained with DAPI and the cells were examined with an Olympus IX51 fluorescence microscope (Olympus, Tokyo, Japan). Images were captured with a CoolPIX 5400 camera (Nikon, Japan).
5
Visualizing Plant Gene Expression
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The plants were monitored daily and analyzed for GFP fluorescence using a handheld UV lamp (7000 μW, Novodirect, Kehl/Rhein, Germany) and for mCherry fluorescence with green light (KL2500 LCD, Schott AG, Mainz, Germany) and a red filter. Pictures were taken with a Nikon Coolpix 5400 camera (Nikon, Düsseldorf, Germany). For microscopic analysis, plant parts were analyzed with a Biorevo BZ-9000 fluorescence microscope (Keyence, Neu-Isenburg, Germany).
6
Viability of Decellularized NPC-Collagen Microspheres
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The morphology of the NPC-collagen microspheres before and after decellularization, the NPC-derived ECM following repopulation with hDFs, and the hDF-collagen microspheres were observed via a Leica phase-contrast light microscope (Leica DMIL, Solms, Germany) and images were captured with a Nikon COOLPIX5400 camera (Nikon, Tokyo, Japan). The viability of the NPCs in the microspheres before and after decellularization, the hDFs seeded in the NPC-derived decellularized matrix and the hDFs seeded in the collagen microspheres was determined by live/dead staining at 6 and 18 days. Cells were incubated for 45 min in the dark with 2 μM calcein acetoxymethylester and 4 μM ethidium homodimer-1 (LIVE/DEAD Viability/Cytotoxicity Kit; Molecular Probes, Invitrogen, Carlsbad, CA, USA), to stain the live and dead cells, respectively. The microspheres were then washed with PBS and examined under a Nikon inverted fluorescent microscope Eclipse TE2000-U equipped with a SPOT FLEX camera (Diagnostic Instruments, Sterling Heights, MI, US).
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