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Fv1000 aom multiphoton system

Manufactured by Olympus
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The FV1000-AOM multiphoton system is a specialized lab equipment designed for high-resolution imaging. It utilizes multiphoton excitation technology to enable deep tissue imaging with low phototoxicity. The system includes an acousto-optic modulator (AOM) for flexible control of the laser beam.

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Lab products found in correlation

Hp ti sa deep see laser system, by Spectra-Physics (2 mentions) Micropore, by 3M (2 mentions) Celltracker green cmfda, by Thermo Fisher Scientific (2 mentions) Volocity software, by PerkinElmer (2 mentions) Epifluorescence microscope, by Olympus (1 mentions) Acti stain 555 phalloidin, by Cytoskeleton (1 mentions) Fluoromount g, by Southern Biotech (1 mentions)

3 protocols using fv1000 aom multiphoton system

1

Intravital Imaging of Neutrophil Migration

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To visualize migration of neutrophils, intravital imaging was performed using FV1000-AOM multiphoton system (Olympus) equipped with a 25x NA1.05 water immersion objective. For two-photon excitation, a Mai-Tai HP Ti:Sa Deep See laser system (Spectra-Physics) was tuned to 900nm for imaging. Neutrophils labeled with CellTracker™ Green CMFDA (Thermo Fisher Scientific Inc.) were intradermally injected into ear skin 2h before in vivo imaging. Mice were anesthetized by intraperitoneal injection of pentobarbital (Nembutal Sodium solution, Oak Pharmaceuticals, Inc., IL) and anesthetic condition was maintained using isoflurane. Hair of the mouse ear was removed using a commercial hair remover (Nair, Princeton, NJ). The anesthetized mice were laid in a ventral recumbent position on a custom-designed stage to expose the dorsal side of the ear pinna for imaging. Micropore™ (3M health care, MN) tapes were placed to immobilize the ear skin. Body temperature of the mice was controlled with heat pad and the ear immersed in a drop of glycerol/saline (40:60 v/v) and covered with a coverslip. Laser injury was induced by focusing the multiphoton laser tuned to 800 nm at a region within the ear dermis for 5 seconds. To track and analyze the movements of the neutrophils, Volocity software (Improvision/Perkin-Elmer, Waltham, MA) and ImageJ (National Institutes of Health, Bethesda, MD) were used.
Arandjelovic S., Perry J.S., Lucas C.D., Penberthy K.K., Kim T.H., Zhou M., Rosen D.A., Chuang T.Y., Bettina A.M., Shankman L.S., Cohen A.H., Gaultier A., Conrads T.P., Kim M., Elliott M.R, & Ravichandran K.S. (2019). A non-canonical role for the engulfment gene ELMO1 in neutrophils that promotes inflammatory arthritis. Nature immunology, 20(2), 141-151.
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2

Intravital Imaging of Neutrophil Migration

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To visualize migration of neutrophils, intravital imaging was performed using FV1000-AOM multiphoton system (Olympus) equipped with a 25x NA1.05 water immersion objective. For two-photon excitation, a Mai-Tai HP Ti:Sa Deep See laser system (Spectra-Physics) was tuned to 900nm for imaging. Neutrophils labeled with CellTracker™ Green CMFDA (Thermo Fisher Scientific Inc.) were intradermally injected into ear skin 2h before in vivo imaging. Mice were anesthetized by intraperitoneal injection of pentobarbital (Nembutal Sodium solution, Oak Pharmaceuticals, Inc., IL) and anesthetic condition was maintained using isoflurane. Hair of the mouse ear was removed using a commercial hair remover (Nair, Princeton, NJ). The anesthetized mice were laid in a ventral recumbent position on a custom-designed stage to expose the dorsal side of the ear pinna for imaging. Micropore™ (3M health care, MN) tapes were placed to immobilize the ear skin. Body temperature of the mice was controlled with heat pad and the ear immersed in a drop of glycerol/saline (40:60 v/v) and covered with a coverslip. Laser injury was induced by focusing the multiphoton laser tuned to 800 nm at a region within the ear dermis for 5 seconds. To track and analyze the movements of the neutrophils, Volocity software (Improvision/Perkin-Elmer, Waltham, MA) and ImageJ (National Institutes of Health, Bethesda, MD) were used.
Arandjelovic S., Perry J.S., Lucas C.D., Penberthy K.K., Kim T.H., Zhou M., Rosen D.A., Chuang T.Y., Bettina A.M., Shankman L.S., Cohen A.H., Gaultier A., Conrads T.P., Kim M., Elliott M.R, & Ravichandran K.S. (2019). A non-canonical role for the engulfment gene ELMO1 in neutrophils that promotes inflammatory arthritis. Nature immunology, 20(2), 141-151.
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3

Visualizing Neutrophil Cytoskeletal Dynamics

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Primary mouse neutrophils were allowed to adhere on an ICAM-1–coated plate in L-15 medium containing glucose (2 mg/ml) and then were stimulated with the appropriate compounds at 37°C. For inhibitor studies, the cells were pretreated with a given inhibitor for 15 min before being stimulated. After 5 min, the cells were washed twice with ice-cold PBS and fixed with 4% paraformaldehyde (PFA) for 15 min at room temperature. The cells were washed with ice-cold PBS and permeabilized with PBS containing 0.1%Triton X-100 for 10 min, which was followed by washing with PBS. Acti-stain 555 phalloidin (150 nM; Cytoskeleton Inc.) was added to the cells and incubated for 20 min, and any excess reagent was washed off with PBS. The cells were visualized, and 10 random images were captured with an epifluorescence microscope (Olympus Inc.) with a 40×, 0.75-NA (numerical aperture) air objective. Random images were selected, and the polarized cells were labeled. Data were pooled, and the number of polarized cells was calculated. For Fig. 7D, the coverslips were mounted on a slide with Fluoromount-G (SouthernBiotech) and imaged with the confocal functionality of a FV1000-AOM multiphoton system (Olympus Inc.) equipped with a 60×, 1.35-NA oil immersion lens (Olympus) using 565-nm excitation.
Surve C.R., To J.Y., Malik S., Kim M, & Smrcka A.V. (2016). Dynamic regulation of neutrophil polarity and migration by the heterotrimeric G protein subunits Gαi-GTP and Gβγ. Science signaling, 9(416), ra22.
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