SYTOX nuclear green stain is impermeable to living cells, but stains nuclei in a syncitium (or otherwise following membrane degradation) (Goonesinghe et al., 2012 (link)). To visualise migration of YSL nuclei relative to the blastoderm margin during epiboly, embryos from heterozygous parents were injected with 1 nl of 0.5 mM Sytox Green fluorescent nucleic acid dye (Invitrogen, United States) into the yolk cell at 3 hpf and then visualised at 4 hpf and 6 hpf under a fluorescence stereomicroscope (Zeiss). Embryos were kept in E3 medium for genotyping verification. Images were captured and processed using a Zeiss AxioCam MR and AxioVision 4.5 software.
Axiocam mr
Manufactured by Zeiss
Sourced in Germany
The AxioCam MR is a high-performance digital camera designed for microscopy applications. It features a fast readout speed, high image quality, and a robust design. The camera is capable of capturing detailed images with a wide range of magnifications and can be integrated with various microscope systems.
Automatically generated - may contain errors
Lab products found in correlation
Axiovision software, by Zeiss (9 mentions)
Axio imager z1, by Zeiss (7 mentions)
Axio observer z1, by Zeiss (6 mentions)
Axiophot microscope, by Zeiss (3 mentions)
Axioskop 2 microscope, by Zeiss (3 mentions)
32 diaminobenzidine tetrahydrochloride dab, by Abcam (2 mentions)
Axioskop 40, by Zeiss (2 mentions)
Axiovision le64, by Zeiss (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Axio imager 2, by Zeiss (2 mentions)
Axiocam hr, by Zeiss (2 mentions)
Incubation system s, by Zeiss (2 mentions)
Plan neofluar 10x 0.3 objective, by Zeiss (2 mentions)
18 well slide, by Ibidi (2 mentions)
Colibri 2, by Zeiss (2 mentions)
Axiovert 200 microscope, by Zeiss (2 mentions)
Lumar v12, by Zeiss (2 mentions)
Ec plan neofluar 20x 0, by Zeiss (2 mentions)
Axio observer z1 inverted microscope, by Zeiss (2 mentions)
Ec plan neofluar 10x 0, by Zeiss (2 mentions)
48 protocols using axiocam mr
1
Visualizing Yolk Syncytial Layer Nuclei Migration
2
Organoid Immunostaining Protocol
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Organoids were fixed in-well with 4% paraformaldehyde (PFA) for 30 min and incubated with 70% ethanol for 1 h at room temperature. Afterwards, crypts were embedded into 2% agarose, dehydrated via a graded ethanol series, embedded into paraffin and sectioned at 5 µm. The sections were processed and stained using standard methods with the following primary antibodies against:
CTSE (1:100, ab36996, Abcam), ECAD (1:200, 610181, BD Biosciences), ITGB1 (1:150, 610467, BD Biosceiences), KI-67 (1:400, #9129, Cell Signaling), LYSC (1:50, sc-27958, Santa Cruz), PKCζ (1:300, sc-216, Santa Cruz), MUC2 (1:50, sc-15334, Santa Cruz). Secondary antibodies were: Alexa Fluor® 488/546 goat anti-rabbit/mouse, Alexa Fluor® 488 donkey anti-goat, Cy3® goat anti-mouse (all 1:200, Invitrogen). Finally, sections were mounted with ProLongTM Gold Antifade with DAPI. Images were taken with the Zeiss AxioObserver Z1 plus ApoTome 2 with an AxioCam MR.
CTSE (1:100, ab36996, Abcam), ECAD (1:200, 610181, BD Biosciences), ITGB1 (1:150, 610467, BD Biosceiences), KI-67 (1:400, #9129, Cell Signaling), LYSC (1:50, sc-27958, Santa Cruz), PKCζ (1:300, sc-216, Santa Cruz), MUC2 (1:50, sc-15334, Santa Cruz). Secondary antibodies were: Alexa Fluor® 488/546 goat anti-rabbit/mouse, Alexa Fluor® 488 donkey anti-goat, Cy3® goat anti-mouse (all 1:200, Invitrogen). Finally, sections were mounted with ProLongTM Gold Antifade with DAPI. Images were taken with the Zeiss AxioObserver Z1 plus ApoTome 2 with an AxioCam MR.
3
Histopathology and IHC Analysis of Organ Tissues
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For histopathology and IHC (Immunohistochemistry), kidneys, liver and spleen were
collected from each group on days 3 and fixed in 4% neutral formalin for 24 h.
Paraffin-embedded tissues were stained with hematoxylin and eosin (H&E) for
histological observation, and analyzed by IHC staining. For IHC, heat-induced
antigen retrieval was performed using 0.01 M sodium citrate buffer in a
microwave for 15 min. After quenching of endogenous peroxidase and blocking in
3% goat serum, tissue was incubated with the primary monoclonal anti-KHV
antibody (P14, Aquatic Diagnostic Ltd.). After three washes with Tris-buffered
saline (TBST), followed by addition of the secondary antibody (goat anti-mouse
IgG) peroxidase activity was detected with
32-diaminobenzidine-tetrahydrochloride (DAB, Abcam). To investigate the
specificity of these reactions, negative and positive controls were used.
Finally, the samples were examined under optical microscopy (AxioCam MR, Carl
Zeiss, Germany).
collected from each group on days 3 and fixed in 4% neutral formalin for 24 h.
Paraffin-embedded tissues were stained with hematoxylin and eosin (H&E) for
histological observation, and analyzed by IHC staining. For IHC, heat-induced
antigen retrieval was performed using 0.01 M sodium citrate buffer in a
microwave for 15 min. After quenching of endogenous peroxidase and blocking in
3% goat serum, tissue was incubated with the primary monoclonal anti-KHV
antibody (P14, Aquatic Diagnostic Ltd.). After three washes with Tris-buffered
saline (TBST), followed by addition of the secondary antibody (goat anti-mouse
IgG) peroxidase activity was detected with
32-diaminobenzidine-tetrahydrochloride (DAB, Abcam). To investigate the
specificity of these reactions, negative and positive controls were used.
Finally, the samples were examined under optical microscopy (AxioCam MR, Carl
Zeiss, Germany).
4
Histopathological Analysis of Koi Herpesvirus
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For histopathology and immunohistochemistry (IHC), kidneys and gills were
collected (n=5) from each group on days 6 and days 9, and fixed in 4%
neutral formalin for 24 h. Paraffin-embedded tissues were stained with
hematoxylin and eosin (H&E) for histological observation, and analyzed by
IHC staining. For IHC, heat-induced antigen retrieval was performed using 0.01 M
sodium citrate buffer in a microwave for 15 min. After quenching of endogenous
peroxidase and blocking in 3% goat serum, tissue was incubated with the
primary monoclonal anti-KHV antibody (P14, Aquatic Diagnostic). After three
washes with Tris-buffered saline (TBST), followed by addition of the secondary
antibody (goat anti-mouse IgG) peroxidase activity was detected with
32-diaminobenzidine-tetrahydrochloride (DAB, Abcam). To investigate the
specificity of these reactions, isotype control as negative control and a known
infected individual as positive control were used. Finally, the samples were
examined under optical microscopy (AxioCam MR, Carl Zeiss, Oberkochen,
Germany).
collected (n=5) from each group on days 6 and days 9, and fixed in 4%
neutral formalin for 24 h. Paraffin-embedded tissues were stained with
hematoxylin and eosin (H&E) for histological observation, and analyzed by
IHC staining. For IHC, heat-induced antigen retrieval was performed using 0.01 M
sodium citrate buffer in a microwave for 15 min. After quenching of endogenous
peroxidase and blocking in 3% goat serum, tissue was incubated with the
primary monoclonal anti-KHV antibody (P14, Aquatic Diagnostic). After three
washes with Tris-buffered saline (TBST), followed by addition of the secondary
antibody (goat anti-mouse IgG) peroxidase activity was detected with
32-diaminobenzidine-tetrahydrochloride (DAB, Abcam). To investigate the
specificity of these reactions, isotype control as negative control and a known
infected individual as positive control were used. Finally, the samples were
examined under optical microscopy (AxioCam MR, Carl Zeiss, Oberkochen,
Germany).
5
Ontogenetic Morphological Changes in Fish
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The hatchlings (n = 20) were sampled daily from hatching to 5 DPH, at 2-day
intervals from 7 to 21 DPH, at 4-day intervals from 25 to 41 DPH, at 6-day intervals from 47 to 83
DPH, and at 30-day intervals from 110 to 170 DPH. The fish were sampled at different stages and
anesthetized with 300 ppm lidocaine-HCl/NaHCO3 at 25oC according to method of
Kang et al. (2005) . Body weight (BW) was measured to the
nearest 0.01 g using an electric balance (AX 200, Shimadzu Corp., Japan), the total length (TL) was
measured to the nearest 0.01 cm using a vernier caliper (CD-20 CP, Mitutoyo, Japan), and growth was
estimated using an exponential growth curve and the calculation of a condition factor.
A mimetic diagram was produced according to the growth for indicate morphological changes. Each
sample of whole fish was fixed in Bouin’s solution for 24 h, washed thoroughly with water, exposed
to decalcification solution for 24 h, washed again, and dehydrated through a graded alcohol series
(70%, 80%, 90%, and 100% ethanol 1 h each). The samples were then cleared with xylene, impregnated
with soft paraffin, impregnated with hard paraffin, embedded, trimmed, and cut (6 μm). The sections
were placed on slides, stained with hematoxylin–eosin, mounted with Canadian balsam, and
examined/photographed under optical microscopy (AxioCam MR, Carl Zeiss, Germany).
intervals from 7 to 21 DPH, at 4-day intervals from 25 to 41 DPH, at 6-day intervals from 47 to 83
DPH, and at 30-day intervals from 110 to 170 DPH. The fish were sampled at different stages and
anesthetized with 300 ppm lidocaine-HCl/NaHCO3 at 25oC according to method of
Kang et al. (2005) . Body weight (BW) was measured to the
nearest 0.01 g using an electric balance (AX 200, Shimadzu Corp., Japan), the total length (TL) was
measured to the nearest 0.01 cm using a vernier caliper (CD-20 CP, Mitutoyo, Japan), and growth was
estimated using an exponential growth curve and the calculation of a condition factor.
A mimetic diagram was produced according to the growth for indicate morphological changes. Each
sample of whole fish was fixed in Bouin’s solution for 24 h, washed thoroughly with water, exposed
to decalcification solution for 24 h, washed again, and dehydrated through a graded alcohol series
(70%, 80%, 90%, and 100% ethanol 1 h each). The samples were then cleared with xylene, impregnated
with soft paraffin, impregnated with hard paraffin, embedded, trimmed, and cut (6 μm). The sections
were placed on slides, stained with hematoxylin–eosin, mounted with Canadian balsam, and
examined/photographed under optical microscopy (AxioCam MR, Carl Zeiss, Germany).
6
Cellular Senescence Imaging Techniques
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Analysis of the SA-β-gal fluorescence assay was carried out on a Zeiss AxioImager Z1 (Carl Zeiss) equipped with an Axiocam MR and using an EC-Plan-Neofluor ×40/1.3 NA objective. Cells displaying > 5 fluorescent granules were considered positive for SA-β-gal activity. Image acquisition of FoxM1, Cyclin B, and Cdc20 immunostaining was performed on a Zeiss AxioImager using a Plan-Apochromat ×63/1.4 NA objective. Prometaphase cells were acquired with 0.24 μM Z-stacks. Image acquisition of H4K20me3, H3K9me3, and Lamin B1 immunostaining was performed on Leica 6000B using a ×40 LD/0.6 NA objective. LAS X software (Leica Microsystems) was used for image acquisition and cells were acquired with 0.44 μM Z-stacks. User-defined fluorescence intensity thresholds were set and used consistently for samples within each experiment. AutoQuant X2 (Media Cybernetics, Rockville, USA) was used for image deconvolution.
7
IVD Tissue Characterization Protocol
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For intact IVDs, post-MRI analysis, a 2 mm wide sagittal tissue segment from the center of the IVD was fixed in periodate lysine paraformaldehyde (PLP) fixative overnight at 4°C. Samples were then washed in PBS and decalcified using Shandon TBD1 Decalcifier solution (ThermoFischer Scientific) over 72 hr at 4°C, changing solution each day. Tissue segments were washed in PBS and placed in 70% ethanol prior to paraffin embedding. Sections of 5 µm were cut and mounted on glass slides. All sections were heated on a hot plate at 55°C for 45 min and deparaffinized and rehydrated. Next, sections were stained with safranin-O/fast green (Sigma-Aldrich, Oakville, ON, Canada) and with antibodies against p16Ink4a and Ki-67 and counter stained using the DAB detection IHC Kit (ab64264, Abcam, Cambridge, MA) following the manufacturer’s instructions. All images were acquired using a Zeiss Axioskop 40 and an AxioCam MR (Zeiss) and processed using AxioVision LE64 software (Zeiss).
8
Zeiss Microscopy Imaging Workflow
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AxioImager 2 (Zeiss) equipped with 20X (0.8 NA) objective and color and monochrome CCD cameras (Axiocam HR and Axiocam MR, Zeiss) was used for routine examination of slides. Select samples were imaged with the laser confocal microscope LSM 700 (Zeiss). Image acquisition was done via the Zen software (Zeiss). Image cropping and image intensity adjustments for figures was done via Photoshop (Adobe).
9
Histological Analysis of Adipose Tissue
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Adipose tissues were excised, fixed in fresh 4% paraformaldehyde (Sigma) in PBS (Gibco; pH 7.4) for 48 h at 4 °C and then embedded with paraffin. Standard hematoxylin and eosin staining was performed on rehydrated 4-micron paraffin sections. Slides were dehydrated and covered with coverslip by resin-based mounting. Tissue sections were visualized by Axiophot microscope equipped with AxioCam MR (Zeiss).
10
Immunostaining Protocol for Cell Fixation
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For immunostaining 0.8×106 cells were seeded onto 16 mm coverslips and allowed to attach for 1 h at RT. Cells were washed briefly with PBS and fixed in 2% (v/v) formaldehyde in PBS for 7.5 min on ice. Cells were permeabilized in PBS, 0.25% (v/v) Triton X-100, 1% (v/v) formaldehyde for additional 7.5 min on ice. Afterwards, cells were washed with PBS and blocked in PBS containing 3% (w/v) BSA for 1 h at RT. Fixed cells were incubated with suitable primary antibody diluted in blocking solution (PBS, 2% BSA, 0.01% Triton X-100, 1.2% Normal Donkey Serum) for 1 h at RT. After two wash steps cells were incubated for an additional hour with suitable secondary antibody at RT. Cells were washed again before DNA counterstaining with DAPI (1 μg/μl in PBS) for 2 min. Cells were washed twice with PBS and mounted using 10 μl Vectashield mounting medium. The coverslip was sealed to the object slide by nail polish. Immunofluorescence was detected using a Zeiss Axiovert 200 epifluorescence microscope equipped with a CDD Camera (AxioCamMR, Zeiss). Images were level-adjusted in FIJI and edited using Adobe Photoshop CS5 and Adobe Illustrator CS5.
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