For atomic forced microscopy (AFM) imaging of liposomal samples, 10 µl of the samples were deposited onto freshly cleaved muscovite Ruby mica sheets (ASTM V1 Grade Ruby Mica from MICAFAB) for 15–20 minutes. Mica sheets are basically negatively charged so samples bind strongly on the mica surface. After 15 min, the samples were dried by using a vacuum dryer. Sometimes the samples were gently washed with 0.5 ml Milli-Q water to remove molecules that were not firmly attached to the mica and the samples were dried as mentioned above. Acoustic alternative current mode AFM was performed using a Pico plus 5500 ILM AFM (Agilent Technologies, USA) with a piezoscanner maximum range of 9 µm. Micro fabricated silicon cantilevers of 225 µm in length with a nominal spring force constant of 21–98 N/m were used from Nano sensors, USA. Cantilever oscillation frequency at 150–300 kHz was tuned into resonance frequency. The images (512 by 512 pixels) were captured with a scan size between 0.5 and 2 µm at a scan speed rate of 0.5lines/S. Images were flattened using Pico view1.4 version software (Agilent Technologies). Image processing and analyzation was done through Pico Image Advanced version software (Agilent Technologies).
Pico image advanced version software
Manufactured by Agilent Technologies
Sourced in United States
Pico Image Advanced version software is a digital image processing and analysis tool developed by Agilent Technologies. It provides advanced features for the visualization, measurement, and analysis of microscopic images. The software supports a variety of image file formats and offers a range of image processing and analysis capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Pico view1.4 version software, by Agilent Technologies (4 mentions)
Pico plus 5500 afm, by Agilent Technologies (4 mentions)
Pico plus 5500 inverted light microscope afm, by Agilent Technologies (2 mentions)
Pico view1.1 version software, by Agilent Technologies (2 mentions)
Microfabricated silicon cantilevers, by Nanosensors (2 mentions)
Silicon cantilevers, by Nanosensors (1 mentions)
Pico plus 5500 ilm afm, by Agilent Technologies (1 mentions)
7 protocols using pico image advanced version software
1
Atomic Force Microscopy of Liposomes
2
Atomic Force Microscopy of Isolated Mitochondria
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Isolated mitochondrial pellet was subsequently processed for AFM as per earlier discussed protocol.51 (link) Contact mode AFM was performed using a Pico plus 5500 AFM (Agilent Technologies, USA) with a piezo scanner of a maximum range of 100μm. Acquired image was processed through Pico Image Advanced version software (Agilent Technologies, USA).
3
Atomic Force Microscopy of Samples
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5 μL of the samples (1 mM) were deposited onto a freshly cleaved muscovite Ruby mica sheet (ASTM V1 Grade Ruby Mica from MICAFAB, Chennai) for 5–10 minutes, and then the sample was dried by using a vacuum dryer. AAC mode AFM was performed using a Pico plus 5500 AFM (Agilent Technologies USA) with a piezo scanner with a maximum range of 9 μm. Micro-fabricated silicon cantilevers of 225 μm in length with a nominal spring force constant of 21–98 N m−1 from Nano sensors were used. Cantilever oscillation frequency was tuned into resonance frequency. The cantilever resonance frequency was 150–300 kHz. The images (256 by 256 pixels) were captured with a scan size between 0.5 and 5 μm at a scan rate of 0.5 lines per s. The images were processed by flatten using Pico view1.4 version software (Agilent Technologies, USA). Image manipulation was done through Pico Image Advanced version software (Agilent Technologies, USA).
4
Atomic Force Microscopy of Nanomaterials
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A total of 5 μL of the samples (1 mM) were deposited onto a freshly cleaved muscovite Ruby mica sheet (ASTM V1 Grade Ruby Mica from MICAFAB, Chennai, India) for 5–10 min and then the sample was dried by using a vacuum dryer. An AAC mode AFM was performed using a Pico plus 5500 AFM (Agilent Technologies Santa Clara, CA, USA), using a piezo scanner, with a maximum range of 9 μm. Microfabricated silicon cantilevers of 225 μm in length with a nominal spring force constant of 21–98 N m−1 from nanosensors were used. The cantilever oscillation frequency was tuned into resonance frequency. The cantilever resonance frequency was 150–300 kHz. The images (256 by 256 pixels) were captured with a scan size between 0.5 and 5 μm at a scan rate of 0.5 lines per s. The images were processed by flatten using Pico view1.4 version software (Agilent Technologies, Santa Clara, CA, USA). Image analysis was done through Pico Image Advanced version software (Agilent Technologies, Santa Clara, CA, USA).
5
Atomic Force Microscopy of Extracellular Vesicles
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For AFM imaging, purified EVs preparations were diluted with filtered PBS (1:100) and a 5 μl aliquot of the diluted sample solution was deposited on freshly cleaved mica sheet followed by incubation at room temperature for 10 min. The dried sample was then gently washed with 200 μl of Milli-Q water to remove salt and loosely bound moieties. AFM experiments were performed in intermittent contact mode (called “tapping” or AAC mode) to minimize tip-induced damage. AAC mode AFM was performed using a Pico plus 5500 inverted light microscope AFM (Agilent Technologies) with a Piezo scanner (maximum range 9 μm). Microfabricated silicon cantilevers 225 μm in length with a nominal spring force constant of 21 to 98 N/m were used (Nano Sensors). Cantilever oscillation frequency was tuned into resonance frequency. The cantilever resonance frequency was 150 to 300 kHz. The images (512 × 512 pixels) were captured with a scan size between 0.5 and 0.8 μm at a scan speed rate of 0.5 lines/s. Images were processed by flatten using Pico view1.1 version software (Agilent Technologies). Image manipulation was done using Pico Image Advanced version software (Agilent Technologies).
6
Atomic Force Microscopy of Biomolecules
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We primarily followed the procedure described previously (Henn et al., 2001 (link)). AFM experiments were performed in intermittent contact mode (called “tapping” or AAC mode) to minimize tip-induced damage. AAC mode AFM was performed using a Pico plus 5500 inverted light microscope AFM (Agilent Technologies) with a Piezo scanner (maximum range 9 µm). Microfabricated silicon cantilevers 225 µm in length with a nominal spring force constant of 21–98 N/m were used (Nano Sensors). Cantilever oscillation frequency was tuned into resonance frequency. The cantilever resonance frequency was 150–300 kHz. The images (512 × 512 pixels) were captured with a scan size between 0.5 and 0.8 µm at a scan speed rate of 0.5 lines/s. Images were processed by flatten using Pico view1.1 version software (Agilent Technologies). Image manipulation was done using Pico Image Advanced version software (Agilent Technologies).
7
Atomic Force Microscopy of Nanostructures
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5 μL of the samples (1 mM) were deposited onto a freshly cleaved muscovite ruby mica sheet (ASTM V1 Grade Ruby Mica from MICAFAB, Chennai) for 5–10 minutes, and then the sample was dried by using a vacuum dryer. AAC mode AFM was performed using a Pico plus 5500 AFM (Agilent Technologies USA) with a piezo scanner with a maximum range of 9 μm. Micro-fabricated silicon cantilevers 225 μm in length with a nominal spring force constant of 21–98 N m−1 from nanosensors were used. The cantilever oscillation frequency was tuned to the resonance frequency. The cantilever resonance frequency was 150–300 kHz. The images (256 by 256 pixels) were captured with a scan size between 0.5 and 5 μm at a scan rate of 0.5 lines per s. The images were processed by flattening using Pico view1.4 version software (Agilent Technologies, USA). Image analysis was done through Pico Image Advanced version software (Agilent Technologies, USA).
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