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Las af 1

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The LAS AF 1.8.2 software is a comprehensive imaging software solution for Leica microscopes. It provides advanced image acquisition, processing, and analysis capabilities for a wide range of applications in scientific research and microscopy.

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Lab products found in correlation

Plan apochromat, by Leica (1 mentions) Dmi6000 cs microscope, by Leica (1 mentions) Dm6000 cfs microscope, by Leica (1 mentions) Sp5 confocal system, by Leica (1 mentions)

Close spelling variants of this product

LAS AF 1.5.1 acquisition software LAS AF v.3.1.0 build 8587 software LAS AF software version 1.6.0 build 1016 LAS AF version 1.6.3 software LAS-AF 1.8.1 Leica LAS AF

4 protocols using las af 1

1

Visualizing Tumor Vasculature in REM-KP Mice

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11-12 week old REM-KPf/fC mice were anesthetized by intraperitoneal injection of ketamine and xylazine (100/20 mg/kg). In order to visualize blood vessels and nuclei, mice were injected retro-orbitally with AlexaFluor 647 anti-mouse CD144 (VE-cadherin) antibody and Hoechst 33342 immediately following anesthesia induction. Pancreatic tumors were removed and placed in HBSS containing 5% FBS and 2mM EDTA. 80-100 micron images in 1024 × 1024 format were acquired with an HCX APO L20x objective on an upright Leica SP5 confocal system using Leica LAS AF 1.8.2 software. Videos were generated using Volocity 3D Image Analysis Software and compressed using Microsoft Video 1 compression.
Fox R.G., Lytle N.K., Jaquish D.V., Park F.D., Ito T., Bajaj J., Koechlein C.S., Zimdahl B., Yano M., Kopp J., Kritzik M., Sicklick J., Sander M., Grandgenett P.M., Hollingsworth M.A., Shibata S., Pizzo D., Valasek M., Sasik R., Scadeng M., Okano H., Kim Y., MacLeod A.R., Lowy A.M, & Reya T. (2016). Image based detection and targeting of therapy resistance in pancreatic adenocarcinoma. Nature, 534(7607), 407-411.
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2

Live-Cell Confocal Imaging Microscopy

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Images were acquired by Leica LAS AF 1.8.2 software with either an inverted Leica SP5 confocal system using a Leica DMI6000CS microscope or an upright Leica SP5 2 confocal system using a Leica DM 6,000 CFS microscope. Using the inverted microscope, images were acquired using a × 10 Leica Plan Apochromat objective with 0.40 numerical aperture for quantification and a × 20 Leica Plan Apochromat objective with 0.70 numerical aperture. Using the upright microscope, images were acquired using an HCX APO L20x objective with a 1.0 numerical aperture for still images and subsequent movies. Imaging of calvarium ranged from 60 to 100 μm. CFP (excitation 458 nm and emission 463–500 nm), GFP (excitation 488 nm, emission 493–556 nm) and DsRed2 (excitation 561 nm, emission 566–650 nm) were excited with an Argon/2 (458, 477, 488, 496 and 514 nm) and Diode pumped solid-state (561 nm) laser, respectively. The power used for dsRed visualization was 8–12% of the appropriate laser. Images were continuously captured in 1,024 × 1,024 or 1,024 × 512 format, with line averaging of 4 (∼10 or 5 s per scan, respectively) for up to 8 h. Multicolour imaging for CFP and GFP were captured sequentially.
Koechlein C.S., Harris J.R., Lee T.K., Weeks J., Fox R.G., Zimdahl B., Ito T., Blevins A., Jung S.H., Chute J.P., Chourasia A., Covert M.W, & Reya T. (2016). High-resolution imaging and computational analysis of haematopoietic cell dynamics in vivo. Nature Communications, 7, 12169.
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3

Visualizing Tumor Vasculature in REM-KP Mice

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11-12 week old REM-KPf/fC mice were anesthetized by intraperitoneal injection of ketamine and xylazine (100/20 mg/kg). In order to visualize blood vessels and nuclei, mice were injected retro-orbitally with AlexaFluor 647 anti-mouse CD144 (VE-cadherin) antibody and Hoechst 33342 immediately following anesthesia induction. Pancreatic tumors were removed and placed in HBSS containing 5% FBS and 2mM EDTA. 80-100 micron images in 1024 × 1024 format were acquired with an HCX APO L20x objective on an upright Leica SP5 confocal system using Leica LAS AF 1.8.2 software. Videos were generated using Volocity 3D Image Analysis Software and compressed using Microsoft Video 1 compression.
Fox R.G., Lytle N.K., Jaquish D.V., Park F.D., Ito T., Bajaj J., Koechlein C.S., Zimdahl B., Yano M., Kopp J., Kritzik M., Sicklick J., Sander M., Grandgenett P.M., Hollingsworth M.A., Shibata S., Pizzo D., Valasek M., Sasik R., Scadeng M., Okano H., Kim Y., MacLeod A.R., Lowy A.M, & Reya T. (2016). Image based detection and targeting of therapy resistance in pancreatic adenocarcinoma. Nature, 534(7607), 407-411.
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In Vivo Imaging of Tumor Vasculature

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Sox10tdTomatoNLS PY230 cells infected with LV-CMV-eGFP were orthotopically transplanted into 6–10 week-old female syngeneic mice. Tumors were allowed to develop until they reached approximately 0.25cm3. For imaging, mice were anesthetized by IP injection of ketamine and xylazine (100/20 mg/kg) and maintained under anesthesia throughout the procedure using 1–2% (vol/vol) isoflurane gas mixed with oxygen. In order to visualize blood vessels and nuclei, mice were injected retro-orbitally with AlexaFluor 647 anti-mouse CD144 (VE-cadherin) antibody and Hoechst 33342 immediately following anesthesia induction. Tumors were exposed by carefully removing hair, skin, and connective tissue while keeping tumor vasculature intact. Mice were then placed inverted on an imaging apparatus, and each tumor was elevated and stabilized on a glass slide to reduce breathing artifacts. 80–150 micron images in 1024 × 1024 format were acquired with an HCX APO L20x objective on an upright Leica SP5 confocal system using Leica LAS AF 1.8.2 software. Videos were generated using Volocity 3D Image Analysis Software and compressed using Microsoft Video 1 compression.
Dravis C., Chung C.Y., Lytle N.K., Herrera-Valdez J., Luna G., Trejo C.L., Reya T, & Wahl G.M. (2018). Epigenetic and transcriptomic profiling of mammary gland development and tumor models disclose regulators of cell state plasticity. Cancer cell, 34(3), 466-482.e6.
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