Vh analyzer

Manufactured by Keyence

Sourced in Japan

The VH analyzer is a lab equipment product from Keyence. It is used for the measurement and analysis of surfaces and textures. The VH analyzer provides accurate and detailed information about the topography and characteristics of a sample's surface.

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Lab products found in correlation

Bz 2, by Keyence (2 mentions) Biozero microscope, by Keyence (1 mentions) Biozero, by Keyence (1 mentions) Dabco, by Merck Group (1 mentions) Biorevo, by Keyence (1 mentions)

8 protocols using vh analyzer

Quantitative Analysis of Abdominal Wound Healing

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After resecting a part of the abdominal wall, wound pictures were taken with a ruler
placed adjacent to the wound, and the wound areas were measured (VH-analyzer: Keyence,
Osaka, Japan). Using histological images stained by H&E and MT stains, the area of
granulation tissue developed between the abdominal muscle wall and the length of
granulation tissue expanded in the lateral direction were morphometrically measured with a
computer-assisted software BZ-II (Keyence). To measure the area of collagenous matrix and
cellular elements in the granulation tissue, images of the representative lesions were
taken using the 20× objective lens in the Bio-Zero microscope (Keyence). In MT-stained
sections, blue-stained collagenous matrix and red-stained cellular elements were
highlighted, binarized, and morphometrically measured using a VH-analyzer, and the
percentages of collagen distribution and cellular element distribution in the granulation
tissue were calculated.

JIMI S., SAPAROV A., KOIZUMI S., MIYAZAKI M, & TAKAGI S. (2021). A novel mouse wound model for scar tissue formation in abdominal muscle wall. The Journal of Veterinary Medical Science, 83(12), 1933-1942.

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Histological Analysis of Liver Necrosis

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Excised liver tissues were fixed immediately with 4% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.4) for histological analysis [20] (link). Sections (4 µm thick) were prepared from paraffin-embedded tissue and subjected to either hematoxylin and eosin (H&E) staining or immunostaining. The level of necrosis (as a percentage of the total area) was estimated by measuring the necrotic area relative to the entire histological section, and an analysis of the necrotic area was performed with a VH analyzer (Keyence, Osaka, Japan). The hemorrhagic area was also determined to quantify the extent of hemorrhage. The results were expressed as a percentage. Sections were also stained for proliferating cell nuclear antigen (PCNA) (Invitrogen, Carlsbad, CA), and the levels measured. Each treatment group comprised five to six mice per time point. The number of PCNA-positive hepatocytes per 1000 hepatocytes was counted in six separate high power fields (×400) per animal. The percentage of PCNA-positive cells was then calculated and the results expressed as a PCNA-labeling index.

Ohkubo H., Ito Y., Minamino T., Eshima K., Kojo K., Okizaki S.I., Hirata M., Shibuya M., Watanabe M, & Majima M. (2014). VEGFR1-Positive Macrophages Facilitate Liver Repair and Sinusoidal Reconstruction after Hepatic Ischemia/Reperfusion Injury. PLoS ONE, 9(8), e105533.

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Preparation and Titration of MERS-CoV Pseudotypes

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At 24 h after the transfection of 293T cells with pKS-MERS-S, pKS-MERS-St7, pKS-MERS-St11, pKS-MERS-St16, or pKS-MERS-St20, the cells were infected with VSV∆G*-G/GFP [9 (link)] at a multiplicity of infection of 0.1 to prepare MERSpv-Swt, MERSpv-St7, MERSpv-St11, MERSpv-St16, or MERSpv-St20, respectively. As a negative control, 293T cells were mock transfected, and the cells were then infected with VSV∆G*-G/GFP to obtain a mock pseudotype. After absorption for 1 h, the inoculum was replaced with culture medium. After a 24-h incubation in a CO₂ incubator, the culture supernatants were collected and stored at −80 °C for later use. For titration of the pseudotype viruses, Vero cells grown in 96-well plates were inoculated with serially diluted pseudoviruses. At 18 hpi, infection with each pseudotype was detected based on GFP expression observed using a fluorescence microscope. The GFP-expressing cells in the photographic images were counted using a VH analyzer (Keyence, Osaka, Japan). Since the area of the photograph is three times smaller than that of a well in the 96-well plate, the number of GFP-expressing cells in each well was calculated. The pseudotype titers were then expressed as focus-forming units (FFU) per mL.

Fukuma A., Tani H., Taniguchi S., Shimojima M., Saijo M, & Fukushi S. (2015). Inability of rat DPP4 to allow MERS-CoV infection revealed by using a VSV pseudotype bearing truncated MERS-CoV spike protein. Archives of Virology, 160(9), 2293-2300.

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Quantifying Muscle Regeneration and Fibrosis

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Centrally nucleated muscle fibers stained with H&E were photographed with an automatic research photomicroscope (AX80; Olympus) and were then counted as regenerating muscle fibers implying fusion of myoblasts^{27 (link)} by VH analyzer (Keyence). Limb interstitial fibrosis detected by Azan staining was morphometrically assessed by the same photomicroscope and software.^{28 (link)–29 (link)}

Masuda H., Tanaka R., Fujimura S., Ishikawa M., Akimaru H., Shizuno T., Sato A., Okada Y., Iida Y., Itoh J., Itoh Y., Kamiguchi H., Kawamoto A, & Asahara T. (2014). Vasculogenic Conditioning of Peripheral Blood Mononuclear Cells Promotes Endothelial Progenitor Cell Expansion and Phenotype Transition of Anti‐Inflammatory Macrophage and T Lymphocyte to Cells With Regenerative Potential. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 3(3), e000743.

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Apoptosis Quantification in Lung and Liver

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Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick endlabeling (TUNEL) was carried out to detect apoptotic DNA breaks in the lung and liver tissues using the TACS 2 TdT-DAB In Situ Apoptosis Detection Kit (Trevigen Inc., Gaithersburg, MD, USA). The slides were counterstained with 1% methyl green. To measure cell death in the lung and liver, TUNEL-positive and -negative alveolar wall cells and hepatocytes were counted until reaching a total number of 1,000 cells in 9–24 nonoverlapping, randomly chosen ×400 fields. Cells were counted using image analysis software from VH Analyzer (Keyence, Osaka, Japan). Data were collected from three individual mice.

Kitamura J., Uemura M., Kurozumi M., Sonobe M., Manabe T., Hiai H., Date H, & Kinoshita K. (2015). Chronic Lung Injury by Constitutive Expression of Activation-Induced Cytidine Deaminase Leads to Focal Mucous Cell Metaplasia and Cancer. PLoS ONE, 10(2), e0117986.

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Quantifying Ischemic Tissue Microvascular Density

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Two weeks after surgery the mice were sacrificed using an overdose of pentobarbital 150 mg/kg/ml (via i.p. administration), and then systemically perfused animals were fixed with 4% paraformaldehyde as described previously [14 (link), 16 (link)]. Ischemic tissues were left in 4% paraformaldehyde overnight at 4°C, and anterior tibial muscles were excised, and embedded into paraffin. For evaluation of infarcted tissue microvascular density (MVD), heat-induced epitope retrieval was performed in deparaffinized tissue sections. Then, sections were incubated overnight with the antibody against anti-rabbit polyclonal CD34 antibody (1:200, clone: EP373Y, Abcam cat. # ab81289). To evaluate pericyte recruitment, Cy3-conjugated monoclonal anti-actin alpha-smooth muscle (α-SMA) antibody (1:200, clone: 1A4, Sigma-Aldrich) was used. The tissue sections for MVD and pericyte recruitment were observed and counted using fluorescence microscopy VH Analyzer (Keyence).

Salybekov A.A., Masuda H., Miyazaki K., Sheng Y., Sato A., Shizuno T., Iida Y., Okada Y, & Asahara T. (2019). Dipeptidyl dipeptidase-4 inhibitor recovered ischemia through an increase in vasculogenic endothelial progenitor cells and regeneration-associated cells in diet-induced obese mice. PLoS ONE, 14(3), e0205477.

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Quantifying Muscle Regeneration Histology

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Total lesion area: Using histological pictures stained by HE and MT stains, the area of granulation tissue developed between the abdominal muscle wall, including fibrosis and regenerative and migrated muscle cells, was morphometrically measured with a computer-assisted-software BZ-II (Keyence).
Fibrosis area: representative lesions in MT stain were taken using 10-time lens in the Bio-Zero (Keyence), and the blue staining collagenous fibers were highlighted, binarized, and measured (VH-analyzer: Keyence, Osaka, Japan), and the percentage of collagen distribution in the granulation tissue was calculated.
Myoblast number: Myogenin-stained granulation tissue was taken using a 10-time lens, and the positive spots in the area of granulation tissue were calculated.
Regenerated myocyte area: Myoglobin-stained granulation tissue was taken using a 10-time lens, and the positive area of regenerative muscles in the granulation area was calculated.
Positive cell number for p-Smad3: Cells with a positive nucleus for p-Smad3 were counted out of the cells in tissue photographed with a 20-time lens. Examined tissues were of the granulation tissue and the stumped muscle (Fig. 7B).

Jimi S., Koizumi S., Sato K., Miyazaki M, & Saparov A. (2021). Collagen-derived dipeptide Pro-Hyp administration accelerates muscle regenerative healing accompanied by less scarring after wounding on the abdominal wall in mice. Scientific Reports, 11, 18750.

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Immunostaining of Microvascular Pericytes

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For alpha‐smooth muscle actin (α‐SMA) staining, paraffin tissue sections, after deparaffinization or the frozen ones, were washed with PBS, blocked with 10% goat serum for 30 minutes at room temperature (RT), then, incubated with Cy3‐conjugated anti‐α‐SMA Ab (Sigma‐Aldrich) and prediluted (1:200 ratio) with 1% BSA/PBS for 2 hours at RT. After washing with PBS, sections were mounted with 1.25% (w/v) DABCO (Sigma‐Aldrich)/90% (v/v) glycerol/10% (v/v) PBS, then observed using a fluorescent microscope (Biorevo; Keyence Corporation, Osaka, Japan). The same protocol, without using the Ab, was performed as the negative control. Also, the negative control for Isolectin B4‐FITC staining was in advance, prepared from the mice without in vivo injecting Isolectin B4‐FITC into the tail vein.
Using software (VH analyzer; Keyence), microvascular densities were evaluated by counting the microvessels stained with Isolectin B4‐FITC. Simultaneously, pericyte recruitment to the vasculartures was evaluated by counting the α‐SMA‐positive microvessels.

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