Viewing chambers were constructed as previously described (41 (link)). Images were recorded on a Nikon Eclipse Ni light microscope fitted with a Hamamatsu C11440 digital camera and Nikon N Plan Apo λ 63×/1.45 NA oil immersion objective. For time-lapse video microscopy, vehicle (DMSO) or RAP-treated schizonts were synchronized by incubation with the reversible PKG inhibitor compound 2 and then washed, mixed in equal proportions, and monitored for egress. Differential inference contrast (DIC) images were taken at 10-s intervals over 45 min, while fluorescence (GFP or mCherry) images were taken every 5 to 10 min to prevent bleaching. Time-lapse videos were analyzed and annotated using Fiji (57 (link)).
C11440
Manufactured by Nikon
The Nikon C11440 is a high-resolution, digital camera module designed for laboratory and scientific applications. It features a 16.2-megapixel CMOS image sensor and can capture images with a resolution of 4936 x 3264 pixels. The device supports a wide range of image data formats, including TIFF, JPEG, and RAW.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse ni light microscope, by Hamamatsu Photonics (3 mentions)
N plan apo λ 63 1.45na, by Nikon (2 mentions)
Mitotracker red cmxros, by Thermo Fisher Scientific (1 mentions)
N plan apo λ 100x 1.45na oil immersion objective, by Nikon (1 mentions)
Eclipse ni e widefield microscope, by Hamamatsu Photonics (1 mentions)
Optima 3000dv, by PerkinElmer (1 mentions)
Chi 661d potentiostat, by CH Instruments (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Ti s l100, by Nikon (1 mentions)
5 protocols using c11440
1
Time-lapse Microscopy of Malaria Parasite Egress
2
Live-cell imaging with fluorescence microscopy
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Viewing chambers were constructed as previously described (38 (link)). Images were recorded on a Nikon Eclipse Ni light microscope fitted with a Hamamatsu C11440 digital camera and Nikon N Plan Apo λ 63×/1.45NA oil immersion objective. For time-lapse video microscopy, differential inference contrast images were taken at 10-s intervals over 30 min, whereas fluorescence (GFP, mTagBFP2, and mCherry) images were taken every 2 min to prevent bleaching. Time-lapse videos were analysed and annotated using Fiji (83 (link)).
3
Live Parasite Microscopy Examination
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Viewing chambers for live parasite microscopic examination were constructed as previously described (7 (link)). All images were recorded on a Nikon Eclipse Ni light microscope fitted with a Hamamatsu C11440 digital camera and Nikon N Plan Apo λ 63×/1.45NA oil immersion objective. For time-lapse video microscopy, differential interference contrast (DIC) images were taken at 10 s intervals over 30 min while fluorescence (mNeon Green) images were taken every 2 min to prevent bleaching. Time-lapse videos were analyzed and annotated using Fiji (44 (link)). For viability staining using the vital mitochondrial dye MitoTracker Red CMXRos (ThermoFisher Scientific; stored as a 10 μM stock in DMSO), the dye was added (20 nM final concentration) to a suspension of schizonts pretreated for 1 h with either DMSO (control, 1% vol/vol) or compound 3j (10 μM). The schizonts were incubated with the dye for 15 min at 37 °C, then washed twice, transferred to a viewing chamber, and observed immediately by dual DIC/fluorescence microscopy.
4
Time-Lapse Microscopy of Malaria Egress
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Time lapse video microscopy of egress was performed as previously described[9 (link)]. Briefly, tightly synchronized schizonts were percoll-enriched and arrested with 1 μM C2 for 4 h. After C2 wash out, DIC and mCherry images were collected every 5 and 25 s, respectively, for 30 min using a Nikon Eclipse Ni-E wide field microscope fitted with a Hamamatsu C11440 digital camera and a Nikon N Plan Apo λ 100x/1.45NA oil immersion objective. For each experiment, videos of the RAP- and DMSO-treated parasites were taken alternately to ensure that possible differences in the rate of egress were not a result of variation in the maturity of the parasite populations. The images were then annotated using Axiovision 3.1 software and exported as AVI movie or TIFF files. Individual egress events were annotated by detailed visual analysis of the movies, and the delay to the time of egress was recorded for each schizont for subsequent statistical analysis. Mean fluorescence intensity values of individual mCherry-expressing schizonts right before and after PVM breakdown were determined from exported raw image files (TIFF format) as described previously[9 (link)] and using the elliptical selection tool and ‘Histogram’ options of ImageJ/Fiji V1.0. DPAP3KO parasites were analyzed to determine the residual background fluorescence derived from the hemozoin.
5
Micromotors for Gastric Delivery
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Template electrochemical deposition of micromotors was carried out with a CHI 661D potentiostat (CH Instruments, Austin, TX). SEM images were obtained with a Phillips XL30 ESEM instrument, using an acceleration potential of 20 kV. The SEM images were taken using fresh micromotor samples. Mapping elemental analysis was carried out using an Oxford EDX attached to the SEM instrument and operated by Inca software. Microscope images and videos were captured by an inverted optical microscope (Nikon Instrument Inc. Ti-S/L100), coupled with a 10× objective, using Hamamatsu digital camera C11440 along with the NIS-Elements AR 3.2 software. The propulsion experiments under physiological temperature were carried out using a Peltier - thermoelectric cooler module (CH-109-1.4-1.5) coupled with a K type thermocouple and a dual digital display PID temperature controller SSR. The size of the gold particle (diameter, nm) was measured by DLS on a Zetasizer Nano ZS (model ZEN3600 from Malvern Instruments). The amount of the released AuNPs retained on the mouse stomach was quantified using an ICP-MS analyzer (PerkinElmer Optima 3000 DV).
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