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Imspector software

Manufactured by Miltenyi Biotec
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Sourced in Germany

The ImSpector Software is a tool for microscope image acquisition, processing, and analysis. It provides a user-friendly interface for controlling various microscope hardware components and capturing high-quality images. The software offers a range of image processing and analysis features to support researchers in their scientific investigations.

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Lab products found in correlation

Trim scope 2, by Miltenyi Biotec (4 mentions) Bx51 wi microscope stand, by Olympus (3 mentions) Xlumplanfl 20x 1.0w, by Olympus (3 mentions) H67080 01, by Hamamatsu Photonics (3 mentions) Axiovert 200 fluorescence microscope, by Zeiss (2 mentions) Illustrator cs2, by Adobe (2 mentions) Plan apochromat oil immersion objective, by Zeiss (2 mentions) Mvx10 zoom body, by Olympus (2 mentions) 5.5 scmos camera, by Olympus (2 mentions) Trimscope, by Miltenyi Biotec (1 mentions) Ultramicroscope 2, by Miltenyi Biotec (1 mentions) Em ccd c9100 camera, by Hamamatsu Photonics (1 mentions) Bx50wi microscope, by Olympus (1 mentions) Trimscope 2 pm system, by Miltenyi Biotec (1 mentions) Matlab script, by MathWorks (1 mentions) Mvplapo 2 objective, by Olympus (1 mentions) H7422 40, by Hamamatsu Photonics (1 mentions) Nir apo, by Nikon (1 mentions) Dmi 6000 tcs sp5, by Leica (1 mentions)

15 protocols using imspector software

1

In Vivo Spinal Cord Imaging Protocol

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The cervical spinal cord was exposed by laminectomy as previously described [38 ]. Briefly, animals were intubated by insertion of a canula in the trachea providing oxygen and isoflurane through a Minivent system (Harvard apparatus). Laminectomy (C2-5) was performed and spinal column fixed at C1 and C6 in a stereotactic frame. Body temperature and heart rate were recorded and stored electronically. Imaging was performed via 2-photon (LaVision TrimScope II microscope, Spectra-Physics laser). Mouse respiration via MiniVent was synchronized to picture acquisition through a triggering device (TrigViFo) reducing imaging artifacts [97 ]. Through an additional toolkit (VivoFollow II) [97 ], this synchronization allowed real-time distortion correction. Tissue displacement was automatically corrected by objective and stage adjustment through a Python script (VivoFollow I) [96 (link)]. Single images, z-stacks and videos were acquired with ImSpector software (LaVision). Raw images were processed with Fiji [82 (link)] and saved as TIF.
Ivan D.C., Berve K.C., Walthert S., Monaco G., Borst K., Bouillet E., Ferreira F., Lee H., Steudler J., Buch T., Prinz M., Engelhardt B, & Locatelli G. (2023). Insulin-like growth factor-1 receptor controls the function of CNS-resident macrophages and their contribution to neuroinflammation. Acta Neuropathologica Communications, 11, 35.
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2

Live Cell Imaging of PTEN-GFP Fusion Proteins

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Live cell imaging was performed as described before [54 (link)] using an inverted Axiovert 200 fluorescence microscope (Zeiss), equipped with 100×/1.4 Plan Apochromat oil-immersion objective (Zeiss), pulsed excitation module (470 nm, 590 nm LEDs), bandpass filters 510–560 nm and gated CCD camera (LaVision, Biotec). Briefly, HeLa cells were seeded onto eight-well chambers pre-coated with a mixture of collagen IV and poly-d-lysine (Ibidi), allowed to attach (24 h) and forward transfected for 24 h with plasmid DNAs encoding PTEN-leader–GFP fusions as indicated. Images were processed using ImSpector software (LaVision, Biotec) and combined in Adobe Illustrator CS2.
Tzani I., Ivanov I.P., Andreev D.E., Dmitriev R.I., Dean K.A., Baranov P.V., Atkins J.F, & Loughran G. (2016). Systematic analysis of the PTEN 5′ leader identifies a major AUU initiated proteoform. Open Biology, 6(5), 150203.
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3

Multiphoton Lifetime Imaging Protocol

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Multiphoton time domain lifetime images were acquired at 20X magnification on a LaVision BioTec TRIMScope, which functions with a temporal bin size of 80ps, according to the manufacturer’s specifications. Time courses were performed by the operator initiating a scan every minute for 10 minutes. Time courses in which refocussing was required were discounted. The TCSPC data files were exported from the La Vision ImSpector Software and lifetime information from individual cells was extracted by fitting the decays from selected areas in the FLIMfit software tool (version 4.7.3) developed at Imperial College London, which has more sophisticated fitting protocols and allows for the fitting of bi-exponential models should it be necessary.
Martin K.J., McGhee E.J., Schwarz J.P., Drysdale M., Brachmann S.M., Stucke V., Sansom O.J, & Anderson K.I. (2018). Accepting from the best donor; analysis of long-lifetime donor fluorescent protein pairings to optimise dynamic FLIM-based FRET experiments. PLoS ONE, 13(1), e0183585.
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4

Multiscale Imaging of Brain and Olfactory Structures

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Brain and snout samples were imaged using a LaVision Ultramicroscope II equipped with infinity-corrected objectives, laser lines OBIS-561nm 100 mW and OBIS-639nm 70 mW, and 595/40 and 680/30 filters for Alexa 555 and Alexa 647 respectively. The olfactory bulbs were imaged with a 4×0,35NA objective, using a laser NA of 0,3 and a step size of 2 µm obtaining images with 1,63 × 1,63 × 2 µm/pixel resolution. The snouts were imaged with a 1,3X objective, adjusting the laser NA to 0,3 and step size of 5 µm obtaining images with a 5 × 5 × 5 µm/pixel resolution. All acquisitions were done with ImSpector software (v.7.0.127.0, Lavision Biotec GmbH). The 3D stacks obtained were analyzed using Bitplane Imaris 9.2 (Oxford instruments).
de Melo G.D., Perraud V., Alvarez F., Vieites-Prado A., Kim S., Kergoat L., Coleon A., Trüeb B.S., Tichit M., Piazza A., Thierry A., Hardy D., Wolff N., Munier S., Koszul R., Simon-Lorière E., Thiel V., Lecuit M., Lledo P.M., Renier N., Larrous F, & Bourhy H. (2023). Neuroinvasion and anosmia are independent phenomena upon infection with SARS-CoV-2 and its variants. Nature Communications, 14, 4485.
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5

High-resolution Imaging of Acute Brain Slices

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High‐resolution videos were obtained using an Olympus microscope with a water immersion objective (20×; NA 0.95). One movie (per condition) was acquired on a TrimscopeTM (LaVision Biotec) using a Ti‐sapphire laser tuned to 820 nm. Images were captured using a Hamamatsu C9100 EM‐CCD camera at 7.65 Hz for a total of 4,000 frames on an area of 450µmX525µm. During data acquisition, acute postnatal mouse brain slices were continuously perfused with oxygenated standard ACSF to approximately 37°C at 1 ml/min. After baseline recordings were obtained, drugs were washed into the bath for 10 min prior to reimaging under the new condition. At the end of each movie, a Z‐stack ± 20 µm (in steps of 1 µm) of the focus plane was acquired for cell body detection. All data collection was done with ImSpector Software (LaVision BioTec).
Pires J., Nelissen R., Mansvelder H.D, & Meredith R.M. (2021). Spontaneous synchronous network activity in the neonatal development of mPFC in mice. Developmental Neurobiology, 81(2), 207-225.
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6

Live Cell Imaging of GFP-VDR Dynamics

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Live cell imaging was performed as described before (77 (link)) using an inverted Axiovert 200 fluorescence microscope (Zeiss), equipped with 100×/1.4 Plan-Apochromat oil-immersion objective (Zeiss), pulsed excitation module (470 nm, 390 nm LEDs), bandpass filters 510–560 nm (EGFP) and 417–477 nm (Hoechst 33342) and gated CCD camera (LaVision BioTec). Briefly, HeLa cells were seeded onto 8-well chambers precoated with a mixture of collagen IV and poly-d-lysine (Ibidi), allowed to attach (24 h) and forward transfected for 24 h with plasmids encoding GFP-VDR fusions as indicated. Prior to live imaging, cells were counterstained with Hoecsht 33342 (1 μm, 30 min). Fluorescence images were collected before (resting) and after stimulation with calcitriol (10−7m, 10 min, 37 °C). Images were exported using ImSpector software (LaVision BioTec) and combined in Adobe Illustrator CS2.
Loughran G., Jungreis I., Tzani I., Power M., Dmitriev R.I., Ivanov I.P., Kellis M, & Atkins J.F. (2018). Stop codon readthrough generates a C-terminally extended variant of the human vitamin D receptor with reduced calcitriol response. The Journal of Biological Chemistry, 293(12), 4434-4444.
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7

Multimodal Imaging of Fibrillar Collagen

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The dissected metatarsus joint area was positioned in close proximity to the surface of a glass-bottom dish (filled with PBS buffer) which was placed above the objective to enable MPI from below. Images with a size of 200 μm × 200 μm were recorded (1,024 × 1,024 pixel, pixel dwell time: 0.7 μs). Z-projections images were calculated in Fiji from the average intensity of (varied number of) images from the z-stack to show the global organization or a certain z-region of the fibrillar collagen network. For larger field-of-view imaging, 2D mosaic image series (8 × 8 images) were taken with an image overlap of 6% using Imspector software (LaVision Biotec). Image stitching was performed using the Autostitcher Plugin of Fiji. The size of the resulting mosaic image was 1,136 μm × 1,136 μm (8,121 × 8,121 pixel). Various regions within the metatarsus region were imaged and the provided images represent typical examples. Sample variability was low.
Vielreicher M., Bozec A., Schett G, & Friedrich O. (2021). Murine Metatarsus Bone and Joint Collagen-I Fiber Morphologies and Networks Studied With SHG Multiphoton Imaging. Frontiers in Bioengineering and Biotechnology, 9, 608383.
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8

Multi-Photon Imaging of Collagen Fibers

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A TriM Scope II multi photon system from LaVision BioTec was used for SHG imaging to visualise anisotropic structures, such as collagen fibres. The microscope setup is a single beam instrument with an upright Olympus BX51 WI microscope stand equipped with high sensitive non-descanned detectors close to the objective lens. The TriM Scope II is fitted with a Coherent Scientific Chameleon Ultra II Ti:Sapphire laser (tuning range 680-1080 nm) and a Coherent Chameleon Compact OPO (automated wavelength extension from 1000 nm to 1600 nm). A 20x IR objective lens (Olympus XLUMPlanFl 20x/1.0W) with a working distance of 2.0 mm was used. SHG signals were detected using TiSa light at 850 nm, a 420/40 band pass filter and a blue-sensitive photomultiplier (Hamamatsu H67080-01). 3D-images were acquired and processed with LaVision BioTec ImSpector Software under Microsoft Windows. Overview images were reconstructed from a sequence of tiled 3D images with Image J.
Langen U.H., Pitulescu M.E., Kim J.M., Enriquez-Gasca R., Sivaraj K.K., Kusumbe A.P., Singh A., Di Russo J., Bixel M.G., Zhou B., Sorokin L., Vaquerizas J.M, & Adams R.H. (2017). Cell-matrix signals specify bone endothelial cells during developmental osteogenesis. Nature cell biology, 19(3), 189-201.
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9

Multi-Photon Imaging of Collagen Fibers

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A TriM Scope II multi photon system from LaVision BioTec was used for SHG imaging to visualise anisotropic structures, such as collagen fibres. The microscope setup is a single beam instrument with an upright Olympus BX51 WI microscope stand equipped with high sensitive non-descanned detectors close to the objective lens. The TriM Scope II is fitted with a Coherent Scientific Chameleon Ultra II Ti:Sapphire laser (tuning range 680-1080 nm) and a Coherent Chameleon Compact OPO (automated wavelength extension from 1000 nm to 1600 nm). A 20x IR objective lens (Olympus XLUMPlanFl 20x/1.0W) with a working distance of 2.0 mm was used. SHG signals were detected using TiSa light at 850 nm, a 420/40 band pass filter and a blue-sensitive photomultiplier (Hamamatsu H67080-01). 3D-images were acquired and processed with LaVision BioTec ImSpector Software under Microsoft Windows. Overview images were reconstructed from a sequence of tiled 3D images with Image J.
Langen U.H., Pitulescu M.E., Kim J.M., Enriquez-Gasca R., Sivaraj K.K., Kusumbe A.P., Singh A., Di Russo J., Bixel M.G., Zhou B., Sorokin L., Vaquerizas J.M, & Adams R.H. (2017). Cell-matrix signals specify bone endothelial cells during developmental osteogenesis. Nature cell biology, 19(3), 189-201.
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10

Light Sheet Microscopy and 3D Imaging

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We separately acquired 16-bit grayscale TIFF images for each channel by light sheet microscopy with the ImSpector software (LaVision Biotec). Tiff stacks were converted (ImarisConverterx64) into Imaris files (.ims) and stitched by Imaris Stitcher. We used Imaris (Bitplane) for 3-D and 2-D image visualization, snapshot creation and movie generation. We performed background subtraction in accordance with the diameter of the respective structures to eliminate unspecific background signals.
All instruments, reagents, software and algorithms used for this study are detailed in Supplementary Table 1.
Mothes R., Pascual-Reguant A., Koehler R., Liebeskind J., Liebheit A., Bauherr S., Philipsen L., Dittmayer C., Laue M., von Manitius R., Elezkurtaj S., Durek P., Heinrich F., Heinz G.A., Guerra G.M., Obermayer B., Meinhardt J., Ihlow J., Radke J., Heppner F.L., Enghard P., Stockmann H., Aschman T., Schneider J., Corman V.M., Sander L.E., Mashreghi M.F., Conrad T., Hocke A.C., Niesner R.A., Radbruch H, & Hauser A.E. (2023). Distinct tissue niches direct lung immunopathology via CCL18 and CCL21 in severe COVID-19. Nature Communications, 14, 791.
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