Pdmi 2

Manufactured by Harvard Apparatus

Sourced in United States, Germany

The PDMI-2 is a dual-channel pressure/differential pressure interface module designed for use with Harvard Apparatus equipment. It serves as an interface between pressure transducers or differential pressure transducers and acquisition systems, providing a means to monitor and record pressure-related data.

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Lab products found in correlation

Mitosox red, by Thermo Fisher Scientific (4 mentions) 510 meta, by Zeiss (3 mentions) Mitosox red, by Harvard Apparatus (2 mentions) Lsm 510, by Zeiss (2 mentions) Te300, by Nikon (1 mentions) Lsm 800, by Zeiss (1 mentions) Poly d lysine, by Merck Group (1 mentions) Hq510lp, by Chroma Technology (1 mentions) Diaphot 300 inverted, by Nikon (1 mentions) 505dcxru, by Chroma Technology (1 mentions) Poly l lysine solution, by Merck Group (1 mentions) 25 mm coverslips, by Thermo Fisher Scientific (1 mentions)

10 protocols using pdmi 2

Quantifying Mitochondrial Calcium Levels

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Qualitative measurement of basal [Ca²⁺]_m were determined as we previously described (Mallilankaraman et al., 2012 (link)). Briefly, cells (5×10⁵/well) grown on 0.2% gelatin-coated coverslips in six-well dishes were loaded with 2 μM rhod-2 AM for 50 min in extracellular medium containing 2 mM Ca²⁺. Coverslips were mounted in an open perfusion microincubator (PDMI-2; Harvard Apparatus) and imaged at 37 °C. All confocal images, obtained at 561 nm excitation using a 63× oil objective, were rapid 100 msec “snapshots” to avoid phototoxicity. All imaging used identical gain and exposure settings that enabled threshold detection of basal Rhod-2 fluorescence in control WT cells. Accordingly, the contribution of low-level diffuse background cytoplasmic Rhod-2 was essentially eliminated. Addition of the mitochondrial uncoupler CCCP (2–10 μM) was added to ensure that punctate fluorescence emanated from mitochondria (Figure S4). Images were analyzed using ImageJ software (NIH) as described (Mallilankaraman et al., 2012 (link)).

Vais H., Mallilankaraman K., Mak D.O., Hoff H., Payne R., Tanis J, & Foskett J.K. (2016). EMRE is a Matrix Ca2+ Sensor that Governs Gatekeeping of the Mitochondrial Ca2+ Uniporter. Cell reports, 14(3), 403-410.

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Temperature-controlled Live-cell Imaging

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Cells were placed in a temperature-controlled chamber (PDMI-2, Harvard Apparatus, Holliston, MA) mounted on the stage of an inverted microscope (Nikon TE300) and recorded using the methods and protocols specified in the Online Supplement.

Hu D., Barajas-Martínez H., Burashnikov A., Panama B.K., Cordeiro J.M, & Antzelevitch C. (2016). Mechanisms Underlying Atrial-Selective Block of Sodium Channels by Wenxin Keli: Experimental and Theoretical Analysis. International journal of cardiology, 207, 326-334.

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Calcium Imaging of Neuronal Responses

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Human primary neurons grown on 50 μg/ml poly-D-lysine (Sigma Aldrich, St Louis)-coated 35-mm glass coverslips were exposed to Tat or TNFα for 48 h. After treatment, the neurons were loaded with 5 μM Fluo-4/AM for 30 min in extracellular medium at room temperature as previously described [37 (link)]. Coverslips were mounted in an open perfusion microincubator (PDMI-2; Harvard Apparatus) at 37 °C and imaged. Confocal images were acquired at 488 nm excitation using a 63× oil objective (LSM 800; Carl Zeiss, Inc.). Images were analyzed and Ca²⁺ fluorescence quantified by using ImageJ (NIH).

Natarajaseenivasan K., Shanmughapriya S., Velusamy P., Sayre M., Garcia A., Gomez N.M, & Langford D. (2020). Inflammation-induced PINCH expression leads to actin depolymerization and mitochondrial mislocalization in neurons. Translational Neurodegeneration, 9, 32.

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Fluorescence Imaging of Labeled Spermatozoa

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Fluorescence imaging was carried out as previously reported (Nishigaki et al., 2004 (link)). Briefly, Fluo-4-labeled spermatozoa were adhered to glass coverslips coated with 50 Pg/ml poly-L-lysine solution (Sigma) and mounted into a micro incubator, PDMI-2 (Harvard Apparatus, Holliston, MA, USA) maintained at 14 °C. Fluorescence images were acquired using a Nikon DIAPHOT 300 inverted microscope with a Nikon Plan Apo 60X objective lens (1.4 NA) and a custom-built stroboscopic illumination system (Nishigaki et al., 2006 (link)) with a Chroma filter set (Ex, HQ470/40x; DC, 505DCXRU; Em, HQ510LP (Chroma Technology)). Images were acquired with a Quantix 57 camera (Photometrics Inc.) under the continuous (stream) acquisition mode.

Beltrán C., Rodríguez-Miranda E., Granados-González G., de De la Torre L.G., Nishigaki T, & Darszon A. (2014). Zn2+ induces hyperpolarization by activation of a K+ channel and increases intracellular Ca2+ and pH in sea urchin spermatozoa. Developmental biology, 394(1), 15-23.

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Mitochondrial ROS Quantification

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To assess levels in reactive oxygen species (ROS), cells were incubated with the mitochondrial oxygen free radical indicator MitoSOX-red (Life Technologies, Carlsbad, CA) for 30 min at 37 °C. Slides were then mounted for confocal imaging in an open perfusion microincubator (PDMI-2; Harvard Apparatus) and images were obtained at 561 nm excitation by using a confocal microscope (810; Carl Zeiss, Oberkochen, Germany). Optical densitometry quantifications were expressed as fluorescence intensity normalized to areas as previously described^{74 (link)}.

Duggan M.R., Mohseni Ahooyi T., Parikh V, & Khalili K. (2021). Neuromodulation of BAG co-chaperones by HIV-1 viral proteins and H2O2: implications for HIV-associated neurological disorders. Cell Death Discovery, 7, 60.

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Quantifying Mitochondrial Superoxide Levels

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Mitochondrial superoxide was measured by using the mitochondrial oxygen free radical indicator MitoSOX Red (molecular probes; Invitrogen) as described previously (70 (link), 71 (link)). Briefly, cells grown on 0.2% gelatin coated glass coverslips were loaded with 5 μM MitoSOX Red for 30 min, and coverslips were mounted in an open perfusion microincubator (PDMI-2; Harvard Apparatus) at 37°C and imaged. Confocal (510 Meta; Carl Zeiss, Inc.) images were obtained at 561 nm excitation by using a 63× oil objective. Images were analyzed, and the mean MitoSOX Red fluorescence was quantified by using Image J software (NIH).

Nemani N., Dong Z., Daw C.C., Madaris T.R., Ramachandran K., Enslow B.T., Rubannelsonkumar C.S., Shanmughapriya S., Mallireddigari V., Maity S., SinghMalla P., Natarajanseenivasan K., Hooper R., Shannon C.E., Tourtellotte W.G., Singh B.B., Reeves B.W., Sharma K., Norton L., Srikantan S., Soboloff J, & Madesh M. (2020). Mitochondrial Pyruvate and Fatty Acid Flux Modulate MICU1-Dependent Control of MCU Activity. Science signaling, 13(628), eaaz6206.

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Measuring Intracellular Calcium Dynamics

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Primary zebrafish cells were grown on Poly-L-lysine coated glass
coverslips and loaded with _mCa²⁺ sensor Rhod-2 (2
μM) for 45 min in extracellular media (ECM-120mM NaCl, 5mM KCl, 1mM
KH₂PO₄, 0.2mM MgCl₂, 0.1mM EGTA, 20mM
HEPES, pH 7.4). Cells were washed and imaged in ECM using Carl Zeiss LSM510
confocal live cell imaging system using the 560 nm excitation (Ex) laser and
580 nm emission (Em) spectra is collected. Ionomycin was added at indicated
time points to increase _cCa²⁺ levels. For
_cCa²⁺ measurements, primary hepatocytes were grown
on 25-mm glass coverslips and transduced with GCaMP6 encoding adenovirus.
Coverslips were mounted in an open perfusion micro-incubator (PDMI-2;
Harvard Apparatus) at 37 C and imaged using the Carl Zeiss LSM510 confocal
live cell imaging system using the Ex-488 nm and Em-510 nm. After 1 min of
baseline recording, vasopressin (100nM) or thapsigargin (2 μM) was
added, and confocal images were obtained every 3 s at 488-nm excitation
using a 40x oil objective. Images were analyzed and quantitated using ZEN
2010 and ImageJ software.

Tomar D., Jaña F., Dong Z., Quinn WJ I.I.I., Jadiya P., Breves S.L., Daw C.C., Srikantan S., Shanmughapriya S., Nemani N., Carvalho E., Tripathi A., Worth A.M., Zhang X., Razmpour R., Seelam A., Rhode S., Mehta A.V., Murray M., Slade D., Ramirez S.H., Mishra P., Gerhard G.S., Caplan J., Norton L., Sharma K., Rajan S., Balciunas D., Wijesinghe D.S., Ahima R.S., Baur J.A, & Madesh M. (2019). Blockade of MCU-Mediated Ca2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation. Cell reports, 26(13), 3709-3725.e7.

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Monitoring Calcium Dynamics in Cardiomyocytes

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Fibroblasts/hiPSCs/cell lineage induced hiPSCs/hiPSCs-CMs with or without MICU1 expression were grown on 0.2% gelatin coated 25 mm glass coverslips. The cells were loaded with 5 μM Fluo-4/AM (30 min) in extracellular medium at room temperature. Coverslips were mounted in an open perfusion microincubator (PDMI-2; Harvard Apparatus) and imaged. Spontaneous Ca²⁺ oscillations were recorded every 3 s (510 Meta; Carl Zeiss, Inc.) at 488 excitations using a ×63 oil objective. Images were analyzed and quantified by using ImageJ (NIH).

Shanmughapriya S., Tomar D., Dong Z., Slovik K.J., Nemani N., Natarajaseenivasan K., Carvalho E., Lu C., Corrigan K., Garikipati V.N., Ibetti J., Rajan S., Barrero C., Chuprun K., Kishore R., Merali S., Tian Y., Yang W, & Madesh M. (2018). FOXD1-dependent MICU1 expression regulates mitochondrial activity and cell differentiation. Nature Communications, 9, 3449.

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Quantifying Mitochondrial Superoxide Levels

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Mitochondrial superoxide was measured by using the mitochondrial oxygen free radical indicator MitoSOX Red (molecular probes; Invitrogen). Briefly, cells grown on 0.2% gelatin coated glass coverslips were loaded with 5 μM MitoSOX Red for 30 min at 37 °C, and coverslips were mounted in an open perfusion microincubator (PDMI-2; Harvard Apparatus) and imaged. Confocal (510 Meta; Carl Zeiss, Inc.) images were obtained at 561 nm excitation by using a ×63 oil objective. Images were analyzed, and the mean MitoSOX Red fluorescence was quantified by using Image J software (NIH).

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Mitochondrial ROS Detection in Transduced Neurons

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Twelve-day old neurons grown on 25 mm coverslips (Fisherbrand, Pittsburg, PA) were transduced with AdNull, Ad-Tat or Ad-U1. Forty-eight hours after treatment, cells were incubated with mitochondrial oxygen free radical indicator MitoSOX red (Life Technologies, Eugene, OR) for 30 min at 37 °C. Coverslips were mounted for confocal imaging in an open perfusion microincubator (PDMI-2; Harvard Apparatus). Images were obtained at 561 nm excitation by using a 63× oil objective of Confocal microscope (810; Carl Zeiss, Inc.).

Torkzaban B., Natarajaseenivasan K., Mohseni Ahooyi T., Shekarabi M., Amini S., Langford T.D, & Khalili K. (2020). The lncRNA LOC102549805 (U1) modulates neurotoxicity of HIV-1 Tat protein. Cell Death & Disease, 11(10), 835.

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