Volocity image acquisition software

Manufactured by PerkinElmer

Sourced in Canada

Volocity is an image acquisition software developed by PerkinElmer. It provides tools for capturing, visualizing, and analyzing images from various microscopy techniques.

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Lab products found in correlation

Lumar v12 fluorescence stereomicroscope, by Zeiss (2 mentions) Axioimager microscope, by Zeiss (2 mentions) Flash 4.0 lt scmos camera, by Hamamatsu Photonics (2 mentions) Volocity quantitation and restoration software modules, by PerkinElmer (2 mentions) Dm4500 microscope, by Leica camera (2 mentions) Ab6584, by Abcam (1 mentions) S2369, by Agilent Technologies (1 mentions) Prolong gold antifade, by Thermo Fisher Scientific (1 mentions) Alexa 647 conjugated goat anti mouse, by Thermo Fisher Scientific (1 mentions) Orca r2 ccd camera, by Hamamatsu Photonics (1 mentions) Ivis spectrum, by PerkinElmer (1 mentions) Living image software, by PerkinElmer (1 mentions) C9100 13 em ccd camera, by Hamamatsu Photonics (1 mentions) Imagepro 7, by Media Cybernetics (1 mentions) Ultraview vox spinning disc confocal microscope, by PerkinElmer (1 mentions) 35 mm glass bottom dishes, by MatTek (1 mentions) Ix81 inverted microscope, by Zeiss (1 mentions) Lsm 510, by Leica camera (1 mentions)

11 protocols using volocity image acquisition software

Chick CAM for Metastatic Modeling

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Use of the chick CAM to model the formation of metastatic colonies and primary tumours is described in detail elsewhere17 (link)18 (link). In brief, fertilized eggs decanted into sterilized weigh boats were allowed to mature ex ovo for 9 days. GFP-expressing cells were injected either i.v. or within the CAM tissue on developmental day 10. At indicated times, the ex ovo chick embryos were placed in a custom intravital imaging chamber46 (link)47 (link). Images of tumour cells in the CAM were acquired using a Lumar V12 fluorescence stereomicroscope (Zeiss) or an Olympus BX61 equipped with a digital camera controlled with Volocity image acquisition software (PerkinElmer).

Sung B.H., Ketova T., Hoshino D., Zijlstra A, & Weaver A.M. (2015). Directional cell movement through tissues is controlled by exosome secretion. Nature Communications, 6, 7164.

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Chick CAM Metastasis Modeling

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Use of the chick CAM to model the formation of metastatic colonies and primary tumors is described in detail elsewhere ^{17 , 18 (link)}. Briefly, fertilized eggs decanted into sterilized weigh boats were allowed to mature ex ovo for 9 days. GFP-expressing cells were injected either IV or within the CAM tissue on developmental day 10. At indicated times the ex ovo chick embryos were placed in a custom intravital imaging chamber ^{46 , 47 (link)}. Images of tumor cells in the CAM were acquired using a Lumar V12 fluorescence stereomicroscope (Zeiss) or an Olympus BX61 equipped with a digital camera controlled with Volocity image acquisition software (PerkinElmer).

Sung B.H., Ketova T., Hoshino D., Zijlstra A, & Weaver A.M. (2015). Directional cell movement through tissues is controlled by exosome secretion. Nature Communications, 6, 7164.

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Quantifying Muscle Fiber Morphology

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To capture single images or z-series stacks, we used an AxioImager microscope (Zeiss), filters for DAPI (excitation/emission 365/445; Zeiss filter set 49), GFP (excitation/emission 470/525; Zeiss filter set 38), Cy3 (excitation/emission 550/605; Zeiss filter set 43 HE), and Cy5 (excitation/emission640/690; Zeiss filter set 50), a Flash 4.0 LT sCMOS camera (Hamamatsu), and Volocity image acquisition software (version 6.3.1; Perkin Elmer). Objectives were ×5 EC Plan-NEOFLUAR NA 0.16, ×10 EC Plan-NEOFLUAR NA 0.3, ×20 Plan-APOCHROMAT NA 0.8, ×40 Plan-APOCHROMAT NA 1.4, and ×63 Plan-APOCHROMAT NA 1.4. To quantitate fluorescence, we used Volocity quantitation and restoration software modules (version 6.3.1; Perkin Elmer), as previously described^{30 (link)}. We measured minimal Feret’s diameter, defined as the minimum distance between parallel tangents^{37 (link)}, by MyHC fiber type in 8 µm frozen sections of TA muscle fibers of LR41;Mbnl1^−/− mice treated with the Clcn1 ASO or invert oligo. Untreated WT served as controls.

Hu N., Kim E., Antoury L, & Wheeler T.M. (2023). Correction of Clcn1 alternative splicing reverses muscle fiber type transition in mice with myotonic dystrophy. Nature Communications, 14, 1956.

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Fluorescence Microscopy Imaging Protocol

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To capture single images or z series stacks, we used an AxioImager microscope (Zeiss), filters for DAPI (excitation/emission 365/445; Zeiss filter set 49), GFP (excitation/emission 470/525; Zeiss filter set 38), Cy3 (excitation/emission 550/605; Zeiss filter set 43 HE), and Cy5 (excitation/emission 640/690; Zeiss filter set 50), a Flash 4.0 LT sCMOS camera (Hamamatsu), a MicroPublisher 3.3 RTV color CCD camera (Q-Imaging), and Volocity image acquisition software (PerkinElmer). Objectives were 5× EC Plan-NEOFLUAR NA 0.16, 10× EC Plan-NEOFLUAR NA 0.3, 20× Plan-APOCHROMAT NA 0.8, 40× Plan-APOCHROMAT NA 1.4, and 63× Plan-APOCHROMAT NA 1.4. To quantitate fluorescence, we used Volocity quantitation and restoration software modules (PerkinElmer), as described previously.¹¹ (link)

Hu N., Kim E., Antoury L., Li J., González-Pérez P., Rutkove S.B, & Wheeler T.M. (2020). Antisense oligonucleotide and adjuvant exercise therapy reverse fatigue in old mice with myotonic dystrophy. Molecular Therapy. Nucleic Acids, 23, 393-405.

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Immunofluorescence Analysis of Fibronectin in Breast Cancer

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Immunofluorescence was performed on representative BCa and adjacent normal tissue from the VUMC cohort. Sections (5µm) were deparaffinized and rehydrated. Antigen retrieval was performed in a steamer either for 30 minutes in pre-warmed modified citrate retrieval buffer (pH 6.0; S2369; Dako) for the total FN or for 12 minutes in Tris-EDTA (TE) buffer (pH 8.0) for ED-A FN. Sections were then blocked in 20% Aquablock/PBS (East Coast Biologics) plus 0.05% Tween-20. The sections being stained for total FN were also treated with an avidin-biotin blocking kit (SP-2001; Vector Laboratories) prior to addition of primary antibody. Primary antibodies were biotinylated rabbit anti-FN (ab6584; Abcam; 1:50) and mouse anti-FN [ED-A] (Clone FN-3E2; 1:200). Detection antibodies were Texas Red conjugated Avidin D (A-1100, Vector Laboratories) and Alexa-647 conjugated goat anti-mouse (Life Technologies). Hoechst 33342 was added to the final incubation step to mark nuclei. Slides were mounted in ProLong Gold Antifade (Invitrogen) and imaged on a BX61 Olympus microscope equipped with a digital camera (Orca ER, Hammamatsu) and controlled with Volocity image acquisition software (PerkinElmer).

Arnold S.A., Loomans H.A., Ketova T., Andl C.D., Clark P.E, & Zijlstra A. (2015). Urinary oncofetal ED-A fibronectin correlates with poor prognosis in patients with bladder cancer. Clinical & experimental metastasis, 33(1), 29-44.

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In Vivo Fluorescence Imaging of Mice

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We removed hair (Nair) and performed all in vivo imaging under general anesthesia consisting of either inhalation isoflurane 1–3% to effect or a cocktail of ketamine 100 mg/kg, xylazine 10 mg/kg, and acepromazine 3 mg/kg by intraperitoneal injection^{6 (link).} We imaged mice using an AxioZoom fluorescence microscope (Zeiss), ×0.5 and ×1.0 objectives, separate filters for GFP (excitation/emission 470/525; Zeiss filter set 38 HE) and DsRed (excitation/emission 550/605; Zeiss filter set 43 HE), an ORCA R2 CCD camera (Hamamatsu), and Volocity image acquisition software (Perkin Elmer). To quantitate fluorescence, we measured DsRed and GFP fluorescence in regions of interest (ROIs) using the Volocity quantitation software module, subtracted background fluorescence, corrected for exposure time, and calculated DsRed/GFP fluorescence ratios. Alternatively, mice were imaged using the IVIS Spectrum (Perkin Elmer) with automatic exposure sequences for excitation/emission 465/520 nm (GFP) and 535/600 nm (DsRed), and quantitated fluorescence in ROIs using Living Image software (Perkin Elmer).

Hu N., Antoury L., Baran T.M., Mitra S., Bennett C.F., Rigo F., Foster T.H, & Wheeler T.M. (2018). Non-invasive monitoring of alternative splicing outcomes to identify candidate therapies for myotonic dystrophy type 1. Nature Communications, 9, 5227.

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Spinning Disc Confocal Microscopy of Cell Imaging

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Cells were grown in 35-mm glass bottom dishes (MatTek) and imaged using an UltraVIEW VoX spinning disc confocal microscope (Perkin Elmer) equipped with a high-sensitivity cooled 14-bit EMCCD C9100-13 camera (Hamamatsu). During imaging, cells were kept in a heated incubation chamber at 37°C with CO₂ (LiveCell, Pathology Devices, Inc.). Volocity image acquisition software was used to capture the images (Perkin Elmer). The track analysis and intensity measurements were done with Image Pro 7.0 (Media Cybernetics).

Singh K., Sharma A., Mir M.C., Drazba J.A., Heston W.D., Magi-Galluzzi C., Hansel D., Rubin B.P., Klein E.A, & Almasan A. (2014). Autophagic flux determines cell death and survival in response to Apo2L/TRAIL (dulanermin). Molecular Cancer, 13, 70.

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Automated Cell Wall Autofluorescence Analysis

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To assess transverse section parameters using autofluorescence of cell walls, pictures were taken with a DM6000 B epifluorescence microscope (Leica) equipped with an ‘A’ filter cube (excitation range: UV; excitation filter: BP 340–380; suppression filter: LP 425). Images were taken using a color Retiga 2000R camera (QIMAGING, Canada) running Volocity image acquisition software (Improvision, UK). When the program is launched, the RGB image is transformed in a gray level (8-bit) image, then a Gaussian filter is applied. The contrast is automatically enhanced and the threshold can be manually adjusted at each step of the analysis to create the selection. Then the selection is enlarged and decreased to smooth it. For cell count, a polar transformation is applied. This allows to draw a line on which local maxima will be counted. The noise tolerance parameter can be adjusted depending on image quality.

Lartaud M., Perin C., Courtois B., Thomas E., Henry S., Bettembourg M., Divol F., Lanau N., Artus F., Bureau C., Verdeil J.L., Sarah G., Guiderdoni E, & Dievart A. (2015). PHIV-RootCell: a supervised image analysis tool for rice root anatomical parameter quantification. Frontiers in Plant Science, 5, 790.

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Imaging and Analyzing Stained Embryos

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Whole mount in situ stained embryos were photographed using a Zeiss Axioplan-2 microscope using Volocity image acquisition software (Improvision, Perkin Elmer Inc.). Embryos processed for fluorescent immunohistochemistry were imaged using either: (1) an Olympus Fluoview FV100 confocal system with an IX81 inverted microscope using Plan-Apo 20X/0.75 NA and Plan-Apo 40X/1.3 NA (oil) objectives, respectively, or (2) a Zeiss LSM780 confocal system with an Observer Z1 inverted microscope and a Plan-Apo 20X/0.8 NA objective. Confocal stacks were analyzed using ImageJ software (Hartig, 2013 (link)).

Subramanian A, & Schilling T.F. (2014). Thrombospondin-4 controls matrix assembly during development and repair of myotendinous junctions. eLife, 3, e02372.

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Bright Field and Immunofluorescence Microscopy

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Bright field and autofluorescence observations were performed using a Leica DM4500 microscope. For autofluorescence, images were taken with the “A” filter cube (excitation range: UV; excitation filter: BP 340–380; suppression filter: LP 425). Immunostained sections were observed with confocal microscopes: Zeiss LSM 510 or Leica SP8. The cell walls were first visualized using autofluorescence [720-nm (chameleon laser) or 405-nm respectively]. The secondary antibody was visualized using a Helium/Neon laser at 543 or 561 nm respectively. Pictures were taken with a color Retiga 2000R camera (QIMAGING, Canada) running Volocity image acquisition software (Improvision, UK).

Henry S., Divol F., Bettembourg M., Bureau C., Guiderdoni E., Périn C, & Diévart A. (2016). Immunoprofiling of Rice Root Cortex Reveals Two Cortical Subdomains. Frontiers in Plant Science, 6, 1139.

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