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Thunder imager 3d

Manufactured by Leica
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The Thunder Imager 3D is a high-performance imaging system designed for 3D cell culture and organoid analysis. It utilizes advanced microscopy techniques to capture high-resolution, 3D images of samples. The system is capable of producing detailed, three-dimensional representations of cellular structures and interactions.

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Lab products found in correlation

Spectra x, by Lumencor (1 mentions) Dfc9000 gtc camera, by Leica (1 mentions) Live dead cell viability assay, by Thermo Fisher Scientific (1 mentions) Cck 8, by Dojindo Laboratories (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Yeasen (1 mentions) Axio scope a1, by Zeiss (1 mentions) Axiocam mrm rev 3, by Zeiss (1 mentions) Dm6 b, by Leica (1 mentions) Ab79935, by Abcam (1 mentions) Gata4, by Santa Cruz Biotechnology (1 mentions) Sc 81707, by Santa Cruz Biotechnology (1 mentions) Sc 25310, by Santa Cruz Biotechnology (1 mentions) Ma5 12960, by Thermo Fisher Scientific (1 mentions) α actinin, by Merck Group (1 mentions) Alexa fluor 488 or 594 secondary antibody, by Thermo Fisher Scientific (1 mentions) Dmi4000, by Leica (1 mentions) Las af software, by Leica (1 mentions) Dfc365fx, by Leica (1 mentions) Rabbit anti iba1, by Abcam (1 mentions) Alexa fluor 488 donkey anti goat igg, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Thunder Imager 3D Tissue epifluorescence microscope THUNDER Imager 3D Cell Culture THUNDER Imager 3D Tissue THUNDER Imager 3D Live Cell and 3D Cell Culture THUNDER Imager 3D Assay THUNDER Imager 3D Cell Culture microscope THUNDER Imager Live Cell & 3D Cell Culture & 3D Assay THUNDER 3D Cell Imager system WD THUNDER Imager Tissue 3D THUNDER Imager 3D Live Cell

9 protocols using thunder imager 3d

1

Fluorescence Imaging with Leica Thunder Imager

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Fluorescence images were recorded with a Leica Thunder Imager 3D based on a DMi8 stand, equipped with a Leica DFC900 GTC sCMOS camera and a LUMENCOR Spectra X as fluorescence excitation source with individually switchable LEDs for specific excitation. DAPI, GFP/Alexa Fluor 488, Cy3 and Alexa647 signals were recorded with a Quad-Band filter cube and an additional emission filter wheel in the emission beam path to avoid channel crosstalk. Image stacks of 13 planes with a step size of 0.42 μm were recorded with a 1.3 NA 63× Glycerol immersion objective at a pixel size of 103 nm. All stacks were deconvolved with Huygens Professional in batch deconvolution mode with standard settings and a SNR of 25.
Image processing was done in Fiji (51 (link)). Images shown are maximum intensity projections of deconvolved stacks. Images of individual cells shown in Supplementary Figure S8G and I were additionally resized by a factor of four without interpolation followed by gaussian filtering with a radius of two pixels.
Müller M., Schauer T., Krause S., Villa R., Thomae A.W, & Becker P.B. (2020). Two-step mechanism for selective incorporation of lncRNA into a chromatin modifier. Nucleic Acids Research, 48(13), 7483-7501.
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2

Evaluating Ciliary Function in Cells and Organoids

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Ciliary beat frequency (CBF) was determined in differentiated ALI cultures and organoids by high-speed video microscopy (HSVM) on a Thunder Imager 3D live Cell using a DFC9000 GTC camera (Leica). ALI cultures were imaged in phase contrast (40× dry objective) and 3D organoids were captured in brightfield (40× dry objective) at 37 °C and 5% CO2. Videos were recorded at 203 frames per second (fps) for 512 frames in total or 404 fps for 1024 frames. For ALI filters, five different locations with moving cilia were selected for each video and beating was determined twice for two seconds. For determination of the wave pattern in organoids, cilia on different cells per condition were observed to validate the presence of an effective and recovery stroke. The same cilia were followed twice for 1–2 s to determine CBF. CBF was performed randomized on coded videos.
Dreyer H.H., van Tuyll van Serooskerken E.S., Rodenburg L.W., Bittermann A.J., Arets H.G., Reuling E.M., Verweij J.W., Haarman E.G., van der Zee D.C., Tytgat S.H., van der Ent C.K., Beekman J.M., Amatngalim G.D, & Lindeboom M.Y. (2023). Airway Epithelial Cultures of Children with Esophageal Atresia as a Model to Study Respiratory Tract Disorders. Children, 10(6), 1020.
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3

Biocompatibility Evaluation of PLGA@ICA-GT@KGN Nanofilm

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To assess the biocompatibility of the PLGA@ICA-GT@KGN nanofilm, BMSCs and chondrocytes suspensions (5 × 105 cells/ml) were implanted separately into culture dishes (serving as a control), PLGA-GT, PLGA@ICA-GT, PLGA-GT@KGN and PLGA@ICA-GT@KGN groups. The BMSCs and chondrocytes-loaded nanofilm were cultured in DMEM supplemented with 10% FBS and 1% antibiotics, with the medium changed every other day.
After 1 and 5 days of in vitro culture, the Live/Dead Cell Viability assay (Invitrogen, USA) was conducted per the manufacturer's instructions [27 (link)]. The samples were observed using a laser confocal microscope (Leica Thunder imager 3D, Germany) to assess BMSC and chondrocyte viability within the scaffold. ImageJ software was used for quantitative analysis of cell viability from live/dead staining images.
Furthermore, to observe the cytoskeleton of BMSCs on the nanofilm, F-actin and nuclei were stained with phalloidin (Yeasen, China) and DAPI (Yeasen, China), respectively, and observed using laser confocal microscopy. The proliferation of BMSCs and chondrocyte were quantitatively evaluated on Days 1 and 5 using a Cell Counting Kit-8 (CCK-8, Dojindo, Japan) by measuring absorbance at 450 nm with a spectrophotometer, following the manufacturer's protocol.
Zhao W., Xu F., Shen Y., Ding Q., Wang Y., Liang L., Dai W, & Chen Y. (2024). Temporal control in shell–core structured nanofilm for tracheal cartilage regeneration: synergistic optimization of anti-inflammation and chondrogenesis. Regenerative Biomaterials, 11, rbae040.
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4

Quantification of Cellular Responses in Brain Lesions

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Images were obtained using the Axio Imager 1 microscope (Zeiss, Oberkochen, Germany). Settings were kept constant for all acquisitions. BrdU and IBA1(or eGFP+): BrdU/IBA1 double positive cells were manually counted in 2 sections per animal located 1.89 & 3.15 mm (mice) and 5 & 5.28 mm (lemur) on the ipsilateral side of the lesion rostral to the epicenter using ImageJ software (National Institutes of Health, USA). Fluoromyelin pictures were acquired with THUNDER Imager 3D (Leica, Wetzlar, Germany; lens ×63).
Poulen G., Aloy E., Bringuier C.M., Mestre-Francés N., Artus E.V., Cardoso M., Perez J.C., Goze-Bac C., Boukhaddaoui H., Lonjon N., Gerber Y.N, & Perrin F.E. (2021). Inhibiting microglia proliferation after spinal cord injury improves recovery in mice and nonhuman primates. Theranostics, 11(18), 8640-8659.
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5

Microscopic Chromosome Imaging Analysis

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Chromosome images were captured using a Zeiss Axio Scope A1 epifluorescence microscope fitted with a high-resolution microscopy camera AxioCam MRm Rev. 3 (Carl Zeiss Ltd. Oberkochen, Germany) and a Leica DM6 B (Leica microsystems, Macquarie park, Australia). Images were analysed using Metasystems Isis FISH Imaging System V 5.5.10 software (Metasystems, Altlussheim, Germany) as well as Thunder Imager 3D (Leica microsystems).
Hill P., Shams F., Burridge C.P., Wapstra E, & Ezaz T. (2021). Differences in Homomorphic Sex Chromosomes Are Associated with Population Divergence in Sex Determination in Carinascincus ocellatus (Scincidae: Lygosominae). Cells, 10(2), 291.
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6

Immunofluorescence Staining of Cardiac Cells

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Cells were fixed in 4% PFA at room temperature for 30 min and washed with PBS twice. Cells were then treated with 10% FBS, 5% BSA and 0.2% Triton X-100 in PBS for 15 min, allowing blocking and permeabilization. Incubation with various primary antibodies was overnight, 4 °C. Following primary antibody incubation and washes in 1X PBS, the cells were incubated with the Alexa-Fluor 488 or 594 secondary antibodies (1:400, Molecular Probes) and nuclei were stained with Dapi (1:5000, Calbiochem). Cells were visualized with a 10×/0.30 or 20×/0.40 (dry lens) objective using inverted microscopes (DMI4000 or THUNDER Imager 3D, Leica Microsystems). The images were captured with a digital camera (DFC365 FX, Leica Microsystems) using LAS-AF software (Leica Microsystems).
Antibodies used in this study are as following: cTNT (MA5-12960, Thermo Fisher Scientific), GATA4 (SC-25310 Santa Cruz), α-MHC (MF20) (14-6503-82, eBioscience), DDR2 (sc-81707, Santa Cruz), CD31 (e-ab-30820, Elabscience), MLC2v (ab79935, Abcam), α-Actinin (A7811, Merck).
Testa G., Russo M., Di Benedetto G., Barbato M., Parisi S., Pirozzi F., Tocchetti C.G., Abete P., Bonaduce D., Russo T, & Passaro F. (2020). Bmi1 inhibitor PTC-209 promotes Chemically-induced Direct Cardiac Reprogramming of cardiac fibroblasts into cardiomyocytes. Scientific Reports, 10, 7129.
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7

Visualizing NLRP3 Inflammasomes in Microglia

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The NLRP3 inflammasomes in microglia were visualized through triple immunofluorescence for NLRP3, the apoptosis-associated speck-like protein containing a CARD (ASC), and IBA-1. The primary antibodies comprised goat anti-NLRP3 (1:500, Millipore, Burlington, MA), mouse anti-ASC (1:1000, Santa Cruz, Dallas, TX), and rabbit anti-IBA-1 (1:1000 Abcam). The secondary antibodies used were donkey anti-goat IgG Alexa Fluor 488 (1:200, Invitrogen), donkey anti-mouse Alexa Fluor 594 (1:200, Invitrogen), and anti-donkey rabbit Alexa Fluor 405 (1:200, Invitrogen). The sections were next examined through 2-μm thick, Z-section analysis using Leica THUNDER 3D Imager. The total number of NLRP3 inflammasomes (i.e., structures positive for both NLRP3 and ASC) per unit area (~216 μm2) of the CA3 subfield was measured using two sections per animal (n = 6/group). Also, the percentage of IBA-1+ microglia containing NLRP3 and ASC positive structures were measured.
Madhu L.N., Kodali M., Attaluri S., Shuai B., Melissari L., Rao X, & Shetty A.K. (2021). Melatonin improves brain function in a model of chronic Gulf War Illness with modulation of oxidative stress, NLRP3 inflammasomes, and BDNF-ERK-CREB pathway in the hippocampus. Redox Biology, 43, 101973.
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8

Microglial Activation and Inflammasome Profiling

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Dual or triple immunofluorescence staining and Z-section analysis in a Nikon confocal microscope or Leica THUNDER 3D Imager were employed for the following measures. 1) Percentages of proinflammatory microglia expressing IBA-1 and CD68 (Madhu et al., 2021 (link)). 2) Number of NLRP3 inflammasomes per 216 μm2 area (i.e., structures co-expressing NLRP3 and ASC) and percentages of microglia with NLRP3 inflammasomes (i.e., cells positive for IBA-1, NLRP3, and ASC) in SSC and the CA1 stratum radiatum (Madhu et al., 2021 (link)). The methods and antibodies employed for these studies are detailed in the supplemental file. Three to six representative sections from each animal (n=5–6/group) were used for these measurements.
Kodali M., Madhu L.N., Reger R.L., Milutinovic B., Upadhya R., Gonzalez J.J., Attaluri S., Shuai B., Gitai D.L., Rao S., Choi J.M., Jung S.Y, & Shetty A.K. (2022). Intranasally Administered Human MSC-derived Extracellular Vesicles Inhibit NLRP3-p38/MAPK Signaling after TBI and Prevent Chronic Brain Dysfunction. Brain, behavior, and immunity, 108, 118-134.
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9

Visualizing Pre- and Postsynaptic Markers

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The presynaptic and postsynaptic markers were visualized through dual immunofluorescence for synaptophysin (Syn) and postsynaptic density protein 95 (PSD95). The primary antibodies comprised anti-rabbit Syn (1:500, synaptic systems, Goettingen, Germany) and anti-goat PSD95 (1:500, Abcam). The secondary antibodies employed were donkey anti-rabbit IgG tagged Alexa Fluor 488 (1:200, Invitrogen) and donkey anti-goat IgG tagged Alexa Fluor 594 (1:200, Invitrogen). For measuring changes in Syn+ and PSD95+ puncta density in the dentate molecular layer (ML) and CA1 stratum radiatum, we employed 0.5-μm thick Z-sections using a Leica THUNDER 3D Imager. The area fraction of Syn+ and PSD95+ puncta were measured from randomly chosen 303 μm2 areas in the dentate ML and the CA1 stratum radiatum (5 areas/region/animal, n = 5–6/group) using Image J.
Madhu L.N., Kodali M., Attaluri S., Shuai B., Melissari L., Rao X, & Shetty A.K. (2021). Melatonin improves brain function in a model of chronic Gulf War Illness with modulation of oxidative stress, NLRP3 inflammasomes, and BDNF-ERK-CREB pathway in the hippocampus. Redox Biology, 43, 101973.
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