Diagnostic instrument rt3 slider

Manufactured by Meyer Instruments

Sourced in United States

The Diagnostic Instrument RT3 Slider is a compact and versatile laboratory equipment piece. It is designed to perform various diagnostic tests and analyses. The core function of this instrument is to provide accurate and reliable results for research and testing applications.

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Lab products found in correlation

Ni u fluorescence microscope, by Nikon (4 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions) Click it plus tunel assay, by Thermo Fisher Scientific (1 mentions) Alexa fluor anti mouse or anti rabbit, by Thermo Fisher Scientific (1 mentions) Mouse monoclonal anti human β actin, by Merck Group (1 mentions) Anti parp, by Santa Cruz Biotechnology (1 mentions) Anti bcl 2, by Cell Signaling Technology (1 mentions) Anti cdk4, by Cell Signaling Technology (1 mentions) Horseradish peroxidase hrp conjugated secondary, by Cell Signaling Technology (1 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Anti ass1, by Santa Cruz Biotechnology (1 mentions) Anti ki 67, by Immunoway (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)

4 protocols using diagnostic instrument rt3 slider

Migration and Invasion Assay Protocol

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For migration assay, 5000 cells were suspended in 250 μl plain medium and seeded in the upper chamber while 750 μl medium containing 10% FBS was placed in the lower chamber. DCA and/or Nic were added to the upper chamber and incubated for 24 h. Cells were fixed with 4% formaldehyde for 30 min and stained with crystal violet for 2 h. Cells on the inner part of the upper chamber were removed. Photos were captured using a Nikon Ni-U fluorescence microscope (Nikon, Tokyo, Japan) equipped with a camera/detector Diagnostic Instrument RT3 Slider (Meyer Instruments, Houston, United States).
For invasion assay, a layer of Matrigel was coated on the upper chamber before adding cells. The number of cells added was 50000 for H28 cells and 500,000 for other cell lines.

Lam S.K., Yan S., Lam J.S., Feng Y., Khan M., Chen C., Ko F.C, & Ho J.C. (2022). Disturbance of the Warburg effect by dichloroacetate and niclosamide suppresses the growth of different sub-types of malignant pleural mesothelioma in vitro and in vivo. Frontiers in Pharmacology, 13, 1020343.

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Quantification of Apoptosis via TUNEL Assay

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TUNEL assay was performed using Click-iT^® Plus TUNEL assay (Invitrogen; Thermo Fisher Scientific, Inc.). De-paraffinization, fixation and permeabilization of formalin-fixed, paraffin-embedded tumour xenograft sections were performed first. Sections were incubated with terminal deoxynucleotidyl transferase (TdT) reaction buffer, and then incubated with TdT buffer containing EdUTP, TdT and TdT enzyme. TUNEL reaction cocktail (Alexa Fluor^® picoyl azide, copper protectant, TUNEL reaction buffer additive and TUNEL reaction buffer) was added to each section. The slides were mounted with Prolong^® Gold antifade reagent (Invitrogen; Thermo Fisher Scientific, Inc.) containing 4′,6-diamidino-2-phenylindole (DAPI). Images were captured using a Nikon Ni-U fluorescence microscope (Nikon, Tokyo, Japan) equipped with a camera/detector Diagnostic Instrument RT3 slider (Meyer Instruments, Houston, TX, USA). Images were captured at ×400 magnification using CFI Plan Fluor DLL 40X objective (Nikon). Images were captured using NIS-Elements Basic Research software (SPOT™ Software 5.0) (Laboratory Imaging Ltd., Prague, Czech Republic).

Lam S.K., Pong U K., Li Y.Y., Xu S., Cheng P.N, & Ho J.C. (2018). Inhibition of ornithine decarboxylase 1 facilitates pegylated arginase treatment in lung adenocarcinoma xenograft models. Oncology Reports, 40(4), 1994-2004.

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Protein Expression Analysis by Western Blot and Immunostaining

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Western blot was performed as previously described [12 (link)]. Specific primary antibodies [mouse monoclonal anti-human β-actin (Sigma-Aldrich), anti-ASS1, anti-OTC, anti-PARP, (Santa Cruz Biotechnology, Inc., Santa Cruz, California, USA), anti-Bcl-2, anti-cleaved caspase-3, anti-cyclin A2, D3, E1, H, anti-CDK4 (Cell Signaling Technology, Danvers, Massachusetts, USA), anti-Ki67 (ImmunoWay Biotechnology Company, Texas, USA), anti-PEG (RevMAb, San Francisco, USA) antibodies] and corresponding horseradish peroxidase (HRP)-conjugated secondary (Cell Signaling Technology) were purchased. An enhanced chemiluminescence (ECL) kit (GE Healthcare) was used to detect protein expression. Beta-actin was selected as reference protein [14 (link)].
Immunohistochemistry and immunofluorescence staining were carried out according to standard protocol. Anti-cytokeratin 1 (Santa Cruz Biotechnology), anti-PEG and corresponding Alexa Fluor anti-mouse or anti-rabbit (Life Technology) antibody were used. The slides were mounted with Prolong® Gold antifade reagent with 4′,6-diamidino-2-phenylindole (DAPI, Life Technologies). Photos were taken with Nikon Ni-U fluorescence microscope (Nikon, Tokyo, Japan) equipped with camera/detector Diagnostic Instrument RT3 Slider (Meyer Instruments, Houston, USA).

Lam S.K., Li Y.Y., Xu S., Leung L.L., U K.P., Zheng Y.F., Cheng P.N, & Ho J.C. (2017). Growth suppressive effect of pegylated arginase in malignant pleural mesothelioma xenografts. Respiratory Research, 18, 80.

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Lung SCC Protein Expression Analysis

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The ASS1, OTC, and ARG2 expression in human lung SCC patient samples was examined by immunohistochemical analysis of lung SCC tumors in paraffin-embedded sections. The research protocol was approved by the Institutional Review Board of the University of Hong Kong/Hospital Authority Hong Kong West Cluster (HKU/HA HKW IRB) (IRB reference number UW 18-019). Samples were collected and analyzed from 12 patients. No consent was required from all subjects. No patient photos were required. Immunohistochemistry staining was carried out according to the standard protocol. Photos were taken using a Nikon Ni-U fluorescence microscope (Nikon, Tokyo, Japan) equipped with a camera/detector Diagnostic Instrument RT3 Slider (Meyer Instruments, Houston, USA).

Lam S.K., Yan S., Xu S., U K.P., Cheng P.N, & Ho J.C. (2019). Endogenous arginase 2 as a potential biomarker for PEGylated arginase 1 treatment in xenograft models of squamous cell lung carcinoma. Oncogenesis, 8(3), 18.

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