Pictures were taken with the light and fluorescence inverted microscope Keyence BZ-9000E, (Keyence Corporation, Higashi-Nakajima, Higashi-Yodogawa-ku, Osaka, 533-8555 1, Japan) using the objectives ×20 (Nicon Plan Apo, 20X/0.75, DIC N2 OFN25 WD 1.0) and ×40 (Nicon Plan Apo, 40X/0.95, DIC M/N2 OFN25 WD 0.14). All pictures were acquired and processed with the Keyence camera and software. The camera is installed in the Keyence BZ-9000E microscope (2/3 inch, 1.5 million pixel monochromer CCD Fotosensor (with LC Filter)) with shooting condition 1360 × 1024 pixel. The acquisition software is delivered with the Keyence BZ-9000E microscope: Program “BZ-II Viewer” (Version: 2.1.00a0.0100.0101.0100.0003). Image acquisition was performed in the following conditions: detector gain +12 dB, bit depth 24, fluorescence filters OP-66834 BZ Filter DAPI-BP (excitation wavelength 360/40 nm, absorption wavelength 460/50 nm), OP-66836 BZ Filter GFP-BP (excitation wavelength 470/40 nm, cold mirror wavelength 505 nm, absorption wavelength 535/50 nm), OP-66838 BZ Filter TexasRed (excitation wavelength 560/40 nm, absorption wavelength 630/60 nm), excitation time 1/15–1/90 s; multifluorescence image acquisitions were performed successively. The obtained pictures were analyzed using the “BZ-II Analyser” (Version 2.1) software, which is delivered with the Keyence BZ-9000E microscope.
Bz 9000e microscope
Manufactured by Keyence
Sourced in Japan
The BZ-9000E is a high-performance fluorescence microscope designed for comprehensive cell and tissue analysis. It features a motorized stage, advanced camera, and user-friendly software interface to capture and analyze detailed fluorescence images. The core function of this product is to provide researchers with a versatile tool for visualizing and quantifying fluorescent samples.
Automatically generated - may contain errors
Lab products found in correlation
Bz 2 analyzer software 1, by Keyence (2 mentions)
Triton x 100, by Thermo Fisher Scientific (2 mentions)
Paraformaldehyde (pfa), by Merck Group (2 mentions)
Bz 9000e, by Keyence (1 mentions)
Dm4000b microscope, by Leica (1 mentions)
Bz2 analyser software, by Keyence (1 mentions)
Rabbit anti phospho histone h3 ser10, by Merck Group (1 mentions)
Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (1 mentions)
Mouse anti flag, by Merck Group (1 mentions)
Alexafluor 546 goat anti rabbit secondary antibodies, by Thermo Fisher Scientific (1 mentions)
Cryostat, by Leica (1 mentions)
Sucrose, by Merck Group (1 mentions)
Bz viewer, by Keyence (1 mentions)
Permafluor, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Illustrator cs6, by Adobe (1 mentions)
Donkey anti rabbit cy5, by Jackson ImmunoResearch (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Axioplan 2 microscope, by Zeiss (1 mentions)
Hoechst, by Merck Group (1 mentions)
11 protocols using bz 9000e microscope
1
Keyence BZ-9000E Fluorescence Microscopy Protocol
2
Quantifying Cardiac Fibrosis by Picrosirius Red
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Harvested LVs were fixed in 4% paraformaldehyde overnight at 4 °C and subsequently embedded in paraffin. Histological sections of 4 µm thickness were stained in picrosirius red solution. Slides were then photographed in 20× magnification in brightfield using a Keyence BZ-9000E microscope (Keyence, Osaka, Japan). The extent of fibrosis was calculated as the area of picrosirius red signal in relation to total tissue area by color thresholding using the Keyence BZ2-Analyser Software.
Left ventricular collagen content was measured using the Abcam total collagen assay kit (ab222942, Abcam, Cambridge, United Kingdom) according to manufacturer’s protocol. Briefly, 10 µg of freshly harvested left ventricles were homogenized in aqua dest. using a Peqlab Tissue Lyser (VWR, Radnor, PA, USA, #000-158) followed by an alkaline hydrolysis to yield free hydroxyproline, which was subsequently oxidized yielding chromophore that was detected using a spectrophotmeter (Thermo Fisher Scientific, Waltham, MA, USA, Type 357, #357-902221T) at OD 560 nm.
Left ventricular collagen content was measured using the Abcam total collagen assay kit (ab222942, Abcam, Cambridge, United Kingdom) according to manufacturer’s protocol. Briefly, 10 µg of freshly harvested left ventricles were homogenized in aqua dest. using a Peqlab Tissue Lyser (VWR, Radnor, PA, USA, #000-158) followed by an alkaline hydrolysis to yield free hydroxyproline, which was subsequently oxidized yielding chromophore that was detected using a spectrophotmeter (Thermo Fisher Scientific, Waltham, MA, USA, Type 357, #357-902221T) at OD 560 nm.
3
Imaging Techniques for Microscopic Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Photomicrographs were acquired using a Leica DM4000B microscope (Leica Microsystems, Wetzlar, Germany) with Diskus software version 4.50.1638-#393 (Hilger, Königswinter, Germany). Immunofluorescence microscopy was conducted using an Eclipse 800E (Nikon, Düsseldorf, Germany) and a Keyence BZ 9000E microscope (Keyence, Neu Isenburg, Germany).
4
Quantification of Cardiac Fibrosis in Aged Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Only mice at the advanced age (553 ± 3 days) were used for this analysis. Cryo-Sects (6 μm thickness) were obtained from excised hearts frozen in liquid nitrogen, fixed in ice-cold acetone, subsequently immersed in Roti®-Histol for 10 min at room temperature, and transferred to water through descending concentrations of ethanol (100%, 96%, 75%). Staining was performed using a 0.1% solution of Sirius Red F3BA in saturated aqueous solution of picric acid for 45 min at 25 °C. Subsequently, slices were rinsed in 1% acetic acid for 2 min. Sections were dehydrated in ascending concentrations of ethanol (75%, 96%, and 100%, each 1 min) and cleared in two stages in Roti®-Histol, 10 min each. Sections were covered with Roti®-Histokitt mounting medium (Carl Roth, Karlsruhe, Germany) and a glass cover slip. After scanning the Picro Sirius Red sections with the Keyence BZ-9000E microscope, images were taken at mid-ventricular level (× 20 magnification), and interstitial fibrosis was quantified as percentage of total tissue area in the field of view. Planimetry was performed using a Keyence BZ2-Analyser software using hybrid cell count algorithm (Keyence, Osaka, Japan).
5
Immunofluorescence Staining of Transfected Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Huh7 and HepG2 cells were seeded on 13 mm coverslips in 12-well plates. Forty-eight hours after transfection with polyethyleneimine, according to the manufacturer’s instructions (as described above), cells were washed with PBS, fixed for 10 min with 3.7% formaldehyde in PBS, subsequently washed with PBS, permeabilized, and blocked with 10% FBS, 0.1% Triton X-100 in PBS at 37 °C. After incubation with the primary antibodies mouse anti-Flag (Sigma-Aldrich, #F1804, 1:100) and rabbit anti-phospho-histone H3 (Ser10) (Sigma-Aldrich, #F1804, 1:250) in blocking solution at RT for 1 h and a brief wash with 0.1% Triton X-100 in PBS, cells were incubated with the Alexa Fluor®488 goat anti-mouse and Alexa Fluor®546 goat anti-rabbit secondary antibodies (1:250, Invitrogen, Carlsbad, CA, USA) and the nuclear dye DAPI in 1% FBS, 0.1% Triton X-100 in PBS at RT for 1 h. After an additional washing step with 0.1% Triton X-100 in PBS, cells were mounted with MOWIOL, and images were obtained with a BZ-9000E microscope (Keyence, Osaka, Japan).
6
Cryopreservation and Histological Analysis of Rat Testes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After perfusion fixation, as described above, the testes were isolated and immersed in 4% PAF overnight at +4 °C. After that, the samples were incubated in 25% sucrose (Sigma, USA) solution in 0.02 M PBS for 36 h for cryoprotection and then frozen in hexane that was cooled to −40 °C with liquid nitrogen. Non-serial sagittal sections were prepared using a cryostat (Leica, Germany), mounted on gelatin-coated glass slides, dried, and stained with hematoxylin and eosin (H&E) using the standard procedures for morphological analyses. The sections were analyzed using a Keyence BZ-9000E microscope (Keyence, Japan) and the BZ Viewer (Keyence, Japan) software.
The randomly chosen, round-shaped, seminiferous tubules with a tubular diameter of 50 were measured on the sections from each animal (n = 10 per group). In each animal, the number of Sertoli cells per tubule was counted in 30 round-shaped tubules at the same stage of spermatogenesis (the elongated spermatid stage). The Sertoli cells were quantified on the basis of their previously reported morphological characteristics and their location in the tubules [54 (link),55 (link)].
The randomly chosen, round-shaped, seminiferous tubules with a tubular diameter of 50 were measured on the sections from each animal (n = 10 per group). In each animal, the number of Sertoli cells per tubule was counted in 30 round-shaped tubules at the same stage of spermatogenesis (the elongated spermatid stage). The Sertoli cells were quantified on the basis of their previously reported morphological characteristics and their location in the tubules [54 (link),55 (link)].
7
Immunofluorescence Microscopy of Chlamydia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunofluorescence cells were seeded in 24 well plates onto coverslips and infected as described above. For microscopy cells were fixed in methanol for 10 minutes or in 4% PFA for 20 minutes, permeabilized for 10 minutes in 0.2% Triton X-100 (Sigma) in PBS and blocked in 5% bovine serum albumin (BSA, Sigma) in PBS. Antibodies used were: rabbit anti-Chlamydia (1∶2.000, Milan Analytica #20-698); Cy5-donkey anti-rabbit (1∶300, Jackson ImmunoResearch); FITC-donkey anti-rabbit (1∶300, Dianova), mouse anti-CPAF clone 100a (1∶200, kind gift from Dr. Guangming Zhong, Department of Microbiology and Immunology, University of Texas Health Science Center at San Antonio, Texas, USA); Cy5-donkey anti-mouse (1∶300, Dianova). Subsequently the samples were stained with Hoechst (1∶15.000, Sigma) for 10 minutes before being mounted in Permafluor (Thermo Fisher). The samples were analyzed with an AxioPlan 2 microscope (Zeiss) using the AxioVision software 4.8.2 (Zeiss) or with a BZ 9000E microscope (Keyence) using the BZ II Analyzer software 1.42 (Keyence). Images were processed and assembled with Adobe Illustrator CS6 (Adobe).
8
Immunocytochemistry of Infected Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Infected neurons were washed twice with 1× ice-cold PBS (Fisher Scientific). The cells were then incubated with 4% paraformaldehyde (Sigma-Aldrich) in 1× PBS for 15 min at room temperature and then washed twice with 1× PBS, permeabilized at room temperature for 20 min with 0.5% Triton X-100 (Fisher Scientific), washed twice with 1× PBS, and blocked for 30 min with 10% goat serum (Gibco) in 1× PBS. Cells were incubated at 4°C overnight in 3% goat serum in 1× PBS with primary antibodies at 1:500 dilution. The next day, the primary antibody solution was removed, and cells were washed three times with 0.05% Tween (Fisher) in 1× PBS (0.05% PBS-T), and incubated with appropriate secondary antibody in 3% goat serum in 1× PBS for 45 min, washed three times with 0.05% PBS-T, and cured overnight using ProLong Anti-Fade Gold with DAPI (Life Technologies). Images were captured with a Keyence BZ9000-E microscope at 20× magnification.
9
Immunocytochemistry of Neuronal Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies for immunocytochemistry were used at dilutions between 1: 100–1: 500 and include the following: NeuN (Millipore ABN78), Doublecortin (CST 4604 S), Nestin (Invitrogen MA1-91657). Neurons were washed twice with 1X ice-cold PBS (Fisher Sci). The cells were then incubated with 4% paraformaldehyde (Sigma-Aldrich) in 1X PBS for 15 min at room temperature and then washed twice with 1X PBS, permeabilized at room temperature for 20 min with 0.5% Triton X-100 (Fisher Sci), washed twice and blocked for 30 min with 10% goat serum (Gibco) in 1X PBS. Cells were incubated at 4 °C overnight in 3% goat serum in 1X PBS with primary antibodies. Next day, primary antibody solution was removed and cells were washed thrice with 0.05% Tween (Fisher) in 1X PBS (0.05% PBS-T), and incubated with appropriate Alexa Fluor® secondary antibody (Lifetech) for 45 min, washed thrice with 0.05% PBS-T, cured overnight with Prolong Anti-Fade Gold with DAPI and imaged. Images were captured with a Keyence BZ9000-E microscope at 40X magnification.
10
GFP-Usp27x Expression Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1205Lu and WM1158 inducible expressing murine or human GFP-Usp27x were stimulated for 24 h with doxycycline and analyzed with a BZ 9000E microscope (Keyence) and photos were processed using the BZ II Analyzer software 1.42 (Keyence).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!