Hematoxylin and Eosin (H&E) staining was performed in 6-μm thick sections of formalin-fixed paraffin-embedded (FFPE) tissues. For immunohistochemistry, FFPE sections were antigen retrieved using citrate buffer (10 mM sodium citrate and 0.05% tween-20 pH 6.0) at 95°C for 15 min. Next, peroxidase blocking was performed for 15 min using methanol containing 0.3% H2O2, followed by Avidin/Biotin (PK6100, Vector Labs) blocking for an additional 15 min at room temperature. The sections were then blocked with 5% goat serum in PBST for 30 min at room temperature and incubated overnight with a primary antibody against TOM20 (Invitrogen, Waltham, MA, USA) in a humidified chamber at 4°C. The sections were subsequently stained with a secondary antibody for 1 h at room temperature and developed using DAB Peroxidase Substrate kit (Vector Labs, Burlingame, CA, USA). Images were acquired using Olympus Provis microscope and processed for noise background equally.
Provis microscope
Manufactured by Olympus
Sourced in Japan, United Kingdom
The Provis microscope is a high-quality optical microscope designed for laboratory use. It features a sturdy, ergonomic design and advanced optics for clear, detailed observations. The Provis microscope is capable of various magnification levels to accommodate a range of sample types and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Nanozoomer 2.0 ht c9600, by Hamamatsu Photonics (2 mentions)
Cool snap camera, by Olympus (2 mentions)
Ndp view2, by Hamamatsu Photonics (2 mentions)
Dcs330 digital camera, by Kodak (2 mentions)
Lsm 710 confocal microscope, by Zeiss (2 mentions)
Avidin biotin pk6100, by Vector Laboratories (1 mentions)
Dab peroxidase substrate kit, by Vector Laboratories (1 mentions)
Dp50 camera, by Olympus (1 mentions)
Ab125212, by Abcam (1 mentions)
2 french fogarty catheter, by Edwards Lifesciences (1 mentions)
Ccd camera, by Olympus (1 mentions)
Bx60 microscope, by Olympus (1 mentions)
Dp 11 digital camera, by Olympus (1 mentions)
F actin, by Thermo Fisher Scientific (1 mentions)
Fv1000 confocal microscope, by Olympus (1 mentions)
Sp5 confocal microscope, by Leica (1 mentions)
Spark 10m, by Tecan (1 mentions)
Moticam 2300 camera, by Olympus (1 mentions)
Ab28364, by Abcam (1 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
18 protocols using provis microscope
1
FFPE Tissue Immunostaining for TOM20
2
Microscopic Analysis of Intestinal Epithelial Maturation
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Microscopic examination was performed using an Olympus PROVIS microscope connected to an Olympus DP50 camera (Olympus, Japan), and the images were evaluated by morphometry using the ImageJ software (National Institutes of Health, Bethesda, MD, USA). The proportion, in percent, of adult-type enterocytes in the villi epithelium was estimated by measuring the length of FcRnneg cells in the proximal SI and the length of non-vacuolated enterocytes in the distal SI, in relation to the total villous length.
One-way ANOVA with multiple comparisons and Tukey’s post-hoc test was performed for analysis of plasma IgG levels during natural development, while Dunnett’s post-hoc test was used for analysis of plasma IgG levels in precociously-induced maturation groups and analysis of the proportion of adult-type epithelium between groups during natural development and in induced maturation groups. The percentage of adult-type epithelium in treatment groups was also plotted for correlation with R2 and Pearson and Spearman test calculated. All statistics were performed using GraphPad Prism version 7 for Mac (GraphPad Software, San Diego, California, USA,www.graphpad.com ). Significance was considered when p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****), or non-significant (ns).
One-way ANOVA with multiple comparisons and Tukey’s post-hoc test was performed for analysis of plasma IgG levels during natural development, while Dunnett’s post-hoc test was used for analysis of plasma IgG levels in precociously-induced maturation groups and analysis of the proportion of adult-type epithelium between groups during natural development and in induced maturation groups. The percentage of adult-type epithelium in treatment groups was also plotted for correlation with R2 and Pearson and Spearman test calculated. All statistics were performed using GraphPad Prism version 7 for Mac (GraphPad Software, San Diego, California, USA,
3
Quantifying Liver Tumor CXCR3-B Expression
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Prior to immunostaining, paraffin-embedded sections were subjected to deparaffinization and antigen retrieval with boiling for 10 min in 0.1 M citrate buffer (with 1 mM EDTA), pH 6.0. The images were acquired using an Olympus Provis microscope. The semi-quantification of DAB OD value was performed using Fiji-ImageJ software. Briefly, liver metastatic tumor nodules were outlined, following with semi-quantification of the staining of CXCR3-B (DAB OD value and the tumor area). The correlation of CXCR3-B and the size of tumor nodules were shown by nonlinear fit line with R-squared value.
4
Carotid Injury and Remodeling in Rats
5
Quantifying Hepatic Fibrosis and Lipid Deposition
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Mouse liver tissues were fixed in 10% paraformaldehyde, embedded in paraffin wax, and thin sectioned (4 μm thickness) before staining. To quantify the hepatic fibrosis area, the prepared sections were stained with picro-sirius red and observed using an Olympus Provis microscope equipped with a CCD camera (Tokyo, Japan), as described previously [35 (link), 36 (link)]. For Oil Red O staining, liver sections were stained with 0.3% Oil Red O in triethyl phosphate in water (60% v/v) for 30 min [37 (link)]. The sections were then observed using a BX-60 microscope with a DP-11 digital camera (Olympus, GmbH, Hamburg, Germany). To ensure data accuracy and reliability, a minimum of 5 chicks was selected for analysis per group, at least 5 sections were studied per chick, and at least 7 randomly selected regions of interest (ROIs) per section were analyzed.
6
Immunofluorescence Analysis of Liver Samples
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The procedures were followed as described previously (Du et al. 2009 (link)). The primary antibodies: PUMA and IRF1 (Cell Signaling Technology), F-actin (Invitrogen), and iNOS (BD Biosciences) were purchased. Slides were viewed with Olympus Provis microscope, and FV1000 confocal microscope (Olympus). Formalin-fixed liver samples were embedded in paraffin, and stained with hematoxylin and eosin (H&E staining) for the assessment of inflammation and tissue damage.
7
Microscopy Techniques for Cellular Imaging
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DAB-revealed sections were imaged using a slide scanner (Nanozoomer 2.0-HT C9600, Hamamatsu, Japan) objective X20 and analyzed with the NDP View2 software (Hamamatsu, Japan). For illustration purposes, bright-field images were exported in tiff format from the nanozoomer images with NDP viewer or captured with a Cool SNAP camera mounted on a provis microscope (Olympus). For immunocytochemical analyses, fluorescence images were acquired on a Leica DM6000 fluorescence microscope using either 40 × /1.25 N.A Plan-apochromat objective (for cell bodies) or a 63 × /1.4 N.A Plan-apochromat (for axons). Confocal images were acquired on a Leica SP5 confocal microscope equipped with an Argon laser (488 nm excitation), a Diode 561 nm and HeNe 633 nm (at a 1024 × 1024 pixel resolution).
8
Transwell Cell Migration Assay
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Approximately 3–5 × 104 cells were plated on inserts (CLS3422-48EA) with media supplemented with 2% FBS containing either DMSO, single drugs, or the combination. Seven hundred and fifty microliters of media containing 7.5% FBS was plated on the lower well (used as an attractant) and cells were allowed to migrate for 16–24 h after which they were washed in PBS, fixed in 4% formaldehyde for 5 min, followed by 20 min in methanol. Cells were finally stained using crystal violet and were imaged using Olympus Provis microscope. To measure the number of migrated cells, the crystal violet was dissolved in methanol and absorbance was measured at 540 nm using Tecan Spark 10 M plate reader instrument.
9
Adipose Tissue Architecture Analysis
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The Olympus Provis microscope (Olympus) was used to image extracted adipose tissue slides. Adipose tissue architecture was assessed by analyzing the tissue morphology of H&E-stained slides. Blood vessels were counted using ImageJ (NIH). Blood vessel lumens were counted using CD31-stained slides, containing five slides per group. CD31-stained slides were used to evaluate 20 random cell surface areas. Beginning from the center of the slide and counting five cells in four directions, ImageJ was used to set the difference in cell areas between the control and treatment groups. In total, 10 random tissue sections were quantified for each sample and means were determined for the aforementioned vessels and vessel lumens.
10
Quantifying Liver Necrosis via H&E Staining
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Formalin-fixed liver samples were embedded in paraffin and evaluated using hematoxylin and eosin (H&E) staining. All the pictures were captured on an Olympus Provis microscope. The necrotic area was assessed quantitatively using Image J software10 (link)32 (link) (National Institutes of Health, USA) as previously described. The ratio of necrotic area/whole area was normalized by the pathologist blinded to our experimental group.
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