Ise module

Manufactured by Roche

The ISE module is a laboratory equipment component used for the measurement and analysis of ion-selective electrodes (ISEs). It is designed to provide precise and reliable data on the concentration of specific ions in a sample. The ISE module's core function is to facilitate the detection and quantification of ionic species, which is crucial for various analytical applications in the fields of chemistry, biochemistry, and environmental science.

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Lab products found in correlation

Cobas 6000, by Roche (3 mentions) Synergy mx, by Agilent Technologies (1 mentions) Cytotoxicity detection kit, by Roche (1 mentions) Cobas 8000, by Roche (1 mentions) Cobas 8000 system, by Roche (1 mentions)

5 protocols using ise module

Quantification of Cellular Potassium

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THP-1 cells were lysed in ultrapure water by five freezing–thaw cycles. The lysates were centrifuged, and K⁺ was quantified by indirect potentiometry using a Cobas 6000 with ISE module (Roche).

Martín-Sánchez F., Martínez-García J.J., Muñoz-García M., Martínez-Villanueva M., Noguera-Velasco J.A., Andreu D., Rivas L, & Pelegrín P. (2017). Lytic cell death induced by melittin bypasses pyroptosis but induces NLRP3 inflammasome activation and IL-1β release. Cell Death & Disease, 8(8), e2984-.

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Measuring Potassium Efflux in BMDMs

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Twelve-well plates with 10⁶ BMDMs per well were stimulated for 4 hours with LPS (1 μg/ml) at 37°C, then washed twice with E-total buffer, and stimulated for 30 min at 37°C with different concentrations of nigericin as indicated in the figure legends or for 2 hours with valinomycin (50 μM) or BB15C5 (50 μM). Then, cells were briefly and quickly washed with nuclease-free water to avoid an osmotic shock and immediately after cells were scraped in 200 μl per well of nuclease-free water followed by three freeze-thaw cycles. Lysates were centrifuged at 16,000g for 10 min at 4°C, and the supernatants were stored at −80°C until K⁺ concentration was quantified by indirect potentiometry using Cobas 6000 with ISE module (Roche).

Tapia-Abellán A., Angosto-Bazarra D., Alarcón-Vila C., Baños M.C., Hafner-Bratkovič I., Oliva B, & Pelegrín P. (2021). Sensing low intracellular potassium by NLRP3 results in a stable open structure that promotes inflammasome activation. Science Advances, 7(38), eabf4468.

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Measuring Cytotoxicity and Caspase-1 Activity

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The presence of LDH in cell supernatants was measured using the Cytotoxicity Detection kit (Roche, Mannheim, Germany), following the manufacturer's instructions. It was expressed as the percentage of the total amount of LDH in the cells. In some control experiments, 25 μM of punicalagin was mixed with a lysate of BMDMs before LDH activity measurement. Caspase-1 activity was measured monitoring the cleavage of the fluorescent substrate z-YVAD-AFC at 400/505 using a Synergy Mx plate reader (BioTek) during 30 min intervals during 6 h. Results are presented as the slope of the RFUs/min. Intracellular K⁺ was quantified from BMDM cell lysates by indirect potentiometry on a Cobas 6000 with ISE module (Roche).

Martín-Sánchez F., Diamond C., Zeitler M., Gomez A.I., Baroja-Mazo A., Bagnall J., Spiller D., White M., Daniels M.J., Mortellaro A., Peñalver M., Paszek P., Steringer J.P., Nickel W., Brough D, & Pelegrín P. (2016). Inflammasome-dependent IL-1β release depends upon membrane permeabilisation. Cell Death and Differentiation, 23(7), 1219-1231.

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Potassium Quantification in Cell Lysates

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Cells were quickly washed and resuspended (by scraping the cells) in nuclease-free water for three freeze-thaw cycles. Lysates were centrifuged at 16,000 g for 10 min at 4 °C^{53 (link)}. Supernatants were taken and K⁺ concentrations were quantified by indirect potentiometric methods using a Cobas 8000 with ISE module (Automatic biochemistry analyzer, Roche).

Wang Y., Zhao N., Jian Y., Liu Y., Zhao L., He L., Liu Q, & Li M. (2022). The pro-inflammatory effect of Staphylokinase contributes to community-associated Staphylococcus aureus pneumonia. Communications Biology, 5, 618.

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Cyst Fluid Analysis Using Thin Prep

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Slide preparation of the cyst fluid was done using the Thin Prep^® Processor (32 ). The Thin Prep^® Processor processes all samples in a similar fashion and deposits cells from a similar sample size within a 20-mm circle on the microscope slide, permitting sample comparisons using uniform methods of cell deposition. Slide staining was done according to the method of Papanicolaou (33 ). Sodium and potassium analysis was done using standard ion-specific electrodes on the ISE Module of the Roche Cobas 8000 system (34 ). Cell counting on the thin-prep and stained slides was done in a semi-automated method, using open-source image analysis software DotCount v1.2 and ImageJ (NIH, Bethesda, MD) (35 ). For each slide, nine non-overlapping images were acquired, which covered over 80% of the sample area. The images were stitched before cell counting was performed. A representative single image is shown in Figure 1 below.
Parameters for the software were similar for all counting sessions and the scores results of the software were used for the analysis. All results of the chemical testing were provided by a spreadsheet from URMC and validated by an independent clinical studies team.

Malik B, & Klock J.C. (2018). Breast Cyst Fluid Analysis Correlations with Speed of Sound Using Transmission Ultrasound. Academic radiology, 26(1), 76-85.

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