[Ca2+]i were measured in Fura-2–loaded cells using an Olympus × 51 microscope (Olympus Centre Valley), with an ORCA-ER camera (Hamamatsu) attached to a Polychrome V (Till Photonics LLC) for HSG cells or acini from mouse SMG. Cells were excited at 340/380 nm with an emission of 510 nm. MetaFluor (Molecular Devices) was used to acquire images and process data.
51 microscope
Manufactured by Olympus
Sourced in Japan
The 1×51 microscope is a high-quality optical instrument designed for laboratory use. It features a monocular eyepiece and a sturdy, stable base. The microscope provides clear, detailed images for a variety of applications.
Automatically generated - may contain errors
Lab products found in correlation
Metafluor, by Molecular Devices (3 mentions)
Orca er camera, by Hamamatsu Photonics (3 mentions)
Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (3 mentions)
Cellsens dimention imaging system, by Olympus (2 mentions)
Dp72 digital camera, by Olympus (2 mentions)
Metafluor imaging system, by Molecular Devices (1 mentions)
Dab peroxidase substrate kit, by Vector Laboratories (1 mentions)
Triplestain ihc kit, by Abcam (1 mentions)
Rabbit anti vwf, by Agilent Technologies (1 mentions)
Rat anti mouse f4 80, by Bio-Rad (1 mentions)
Dp72 camera, by Olympus (1 mentions)
Cellsens standard, by Olympus (1 mentions)
Cellsens dimension imaging system, by Olympus (1 mentions)
11 protocols using 51 microscope
1
Intracellular Calcium Measurement in Cells
2
Caspase-3 Activity Assay in HSG Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Active caspase-3 was detected in HSG cells and isolated submandibular acini as described earlier6 (link) using the CellEvent Caspase-3/7 Green Detection Reagent (Thermo Fisher Scientific) with an absorption/emission maximum of 502/530 nm. HSG cells and isolated submandibular acini were plated on glass coverslips and incubated with the reagent at 1 μM concentration for 30 min. The reagent was detected using an Olympus × 51 microscope (Olympus), with an ORCA-ER camera (Hamamatsu) attached to a Polychrome V (Till Photonics LLC). MetaFluor (Molecular Devices) was used to acquire images and process data as described above. Apoptotic cells with activated caspase3/7 have bright green nuclei, whereas cells without activated caspase-3 have minimal fluorescence signal.
3
Fura-2 Calcium Imaging Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded on gelatin-coated glass coverslips at a density of 5000 cells/cm2 at 24 h before the experiments. Cells were starved in DMEM 5% FBS for at least 2 h before the experiments. Cells were next loaded (45 s at 37 °C) with 2 μM Fura-2 AM, for ratiometric cytosolic calcium concentration ([Ca2+]i) measurements. During experiments, cells were maintained in standard extracellular solution of the following composition: 154 mM NaCl, 4 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 5 mM HEPES, 5.5 mM glucose. Solution pH was adjusted to 7.35 using NaOH. 4 h before experiments cells were starved in DMEM 2% FBS. Fluorescence measurements were made using a Polychrome V spectrofluorimeter (TILL Photonics, Munich, Germany) attached to an Olympus ×51 microscope (Olympus, Tokyo, Japan) and Metafluor Imaging System (Molecular Devices, Sunnyvale, CA, USA). [Ca2+]i was measured using ratiometric probe Fura-2-AM and quantified according to Fiorio Pla et al. [50 (link)].
4
Measuring Intracellular Calcium in Salivary Gland Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
[Ca2+]i in HSG cells or acini from mouse SMG were measured in Fura-2–loaded cells using an Olympus × 51 microscope (Olympus Centre Valley), with an ORCA-ER camera (Hamamatsu) attached to a Polychrome V (Till Photonics LLC). Cells were excited at 340/380 nm with an emission of 510 nm. MetaFluor (Molecular Devices) was used to acquire images and process data.
5
EdU incorporation assay for cell proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
EdU incorporation assay was determined by using a Cell-Light EdU Apollo 488 In Vitro Imaging Kit (Ruibo Company, Shanghai, China) according to the manufacturer’s instruction. The images were acquired with an Olympus ×51 microscope (Olympus), and EdU-positive cells were counted.
6
Quantifying EpCAM and CD133 Expressions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The frozen tissues were sectioned, and slides were fixed in cold acetone (−20°C) for 2 minutes. After fixing, the slides were washed with PBS-T, and blocked with 10% FBS for 30 minutes at room temperature. The slides were washed again and exposed to 1:200 dilution of FITC-conjugated antibodies (anti-EpCAM and anti-CD133) to identify the expression of EpCAM and CD133 within the cancerous tissue treatments of Adr/GNRs@PMs-antiEpCAM and Adr/GNRs@PMs before and after laser exposure. All digital images were acquired with the Olympus 1×51 microscope at 40× magnification using the Olympus DP72 digital camera via the cellSens Dimention imaging system (Olympus Corporation) and stored as JPG data files, with fixed resolutions of 200 pixels/inch. FITC-positive staining was examined under a fluorescence microscope at 200× magnification. A computer image analysis was performed to quantify the expressions of EpCAM and CD133 in Adr/GNRs@PMs-antiEpCAM and Adr/GNRs@PMs before and after laser. The acquired color images from the immuno-histochemical staining were defined at a standard threshold according to the software specification. The computer program then quantified the threshold area represented by color images. CD133 and EpCAM protein expressions were defined by the percentages of threshold area in acquired color images.
7
Immunohistochemical Staining of Tissue Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical staining was performed on the paraffin-embedded tissue sections. Endogenous peroxidase was blocked with 3% hydrogen peroxide, and then with 5% animal serum for 30 min to block non-specific reactions. These tissue sections were incubated with primary antibodies (see antibody list in Additional file 1 ). Tissue sections were incubated with horseradish peroxidase-conjugated secondary antibodies (1, 300–400 dilutions with PBS) for 2 h at room temperature, and then incubated with peroxidase substrate DAB kit (Vector Laboratories, Inc., Burlingame, CA) to develop brown color. The counterstaining was performed using hematoxylin or methyl green. A negative controls without primary antibodies was included in each run. Triple staining was performed on human sample using a Triplestain IHC Kit (Abcam, ab183286, Cambridge, MA) according to the provided factory instructions. Digital images were acquired with the Olympus 1 × 51 microscope (Olympus, Pittsburgh, PA) at 10× magnification using the Olympus DP72 digital camera and the length of scratch-wound was measured via the cellSens Dimention imaging system.
8
Scratch-Wound Healing Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For scratch-wound healing assays, cells were seeded in 24-well plates with regular media. After a 24-h acclimatization period, the cells were washed with PBS and cultured for 48 h in FFA or BSA media. Following the 5-day incubation, cells were washed with PBS. Using the tip of a sterile 10 uL pipette, a single scratch was made on the cell surface within each well. Following the scratch for 24 h and 48 h, digital images were acquired with the Olympus 1 × 51 microscope (Olympus, Pittsburgh, PA) at 10× magnification using the Olympus DP72 digital camera. Three standardized bright-field images were recorded for each scratch at the 0, 24, and 48-h time points. The length of scratch-wound was measured via the cellSens Dimention imaging system.
9
Quantification of FGF19, FGFR4 and EpCAM in Liver Diseases
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A computer image analysis was performed to quantify the expressions of FGF19, FGFR4 and EpCAM in the 24 samples diagnosed with ST, NASH, CR, HCC, and in the paired samples of peritumoral liver tissues. The imaging fields were chosen randomly from various section levels to ensure objectivity of sampling. Five imaging fields were scanned for each specimen sample. All digital images were acquired with the Olympus 1 × 51 microscope at 40× magnification using the Olympus DP72 digital camera via the cellSens Dimention imaging system (Olympus, Pittsburgh, PA) and stored as JPG data files, with fixed resolutions of 200 pixels/inch. The acquired color images from the immunohistochemical staining were defined at a standard threshold according to the software specification. The computer program then quantified the threshold area represented by color images. FGF19, FGFR4 and EpCAM protein expressions were defined by the averages of threshold area in acquired color images.
10
Immunostaining of Paraffin-Embedded Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin embedded tissue sections were used for immunostaining with rat anti-mouse F4/80 (AbD Serotec), rabbit anti-vWF (Dako), rat anti-mouse ICAM (BioLegend), rat anti-mouse CD34 (Cedarline), and mouse anti-MSP1 (R&D Systems). AEC and DAB kits were obtained from Vector Laboratory. Images were acquired with an Olympus 1×51 microscope with an Olympus DP 72 camera. Quantification of positive staining was performed using ImageJ Fiji version and CellSens Standard (Olympus) software.
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!