synthesize nanoparticles using the thermal decomposition method as reported (Sun et al #####%%%%% output "nanoparticles" are used in line6
{"action": "synthesize", "output": "nanoparticles", "reagent": [""]}
Cool the black-colored mixture at room temperature by removing the heat source, further precipitated using ethanol, .
{"action": "cool", "output": "", "reagent": ["ethanol"], "temperature": ["room temperature"]}
Discard the brown supernatant . #####%%%%% output "the brown supernatant" are not used
{"action": "remove", "output": "the brown supernatant", "reagent": ["nanoparticles"]}
wash the nanoparticles once more with ethanol to remove leftover nonmagnetic product in the solution #####%%%%% output "leftover nonmagnetic product" are not used
{"action": "rinse", "output": "leftover nonmagnetic product", "reagent": ["ethanol"]}
Note: Further washing with ethanol results in the etching of the oleic acid layer from the SPMNPs (this step might be useful depending on the hydrophobic to hydrophilic reaction step). #####%%%%% output "hydrophilic reaction step" are not used
{"action": "note", "output": "hydrophilic reaction step", "reagent": ["ethanol"]}
After isolation of black powders through centrifugation, add 1 mL of water, , . #####%%%%% output "black powders" are used in line28
{"action": "add", "output": "black powders", "reagent": ["water"]}
then adjust to pH 7 #####%%%%% output "TGA" are used in line16
{"action": "adjust", "output": "TGA", "time": [""]}
Further determine the SPMNPs concentration using TGA respectively. #####%%%%% output "FIASH-EDT2" are used in line18
{"action": "determine", "output": "FIASH-EDT2", "reagent": ["TGA"]}
Perform Fluorescein conjugated DMSA functionalized SPMNPs by mixing 25-fold molar excess of FIASH-EDT2-Maleimide over molar amount of SPMNPs to be coupled.
{"action": "perform", "output": "", "reagent": ["SPMNPs", "FIASH-EDT2", "Maleimide"]}
Perform the reaction overnight at room temperature. #####%%%%% output "the reaction" are not used
{"action": "perform", "output": "the reaction", "temperature": ["room temperature"]}
Remove unbound (fluorescein) by extensive washing in the presence of a magnetic field. #####%%%%% output "PBS" are used in line24
{"action": "remove", "output": "PBS", "reagent": ["unbound (fluorescein)"], "device": ["a magnetic field"]}
Magnetically purified fluorescein conjugated SPMNPs are resuspended in PBS for further cellular uptake analysis. #####%%%%% output "further cellular uptake analysis" are not used
{"action": "rinse", "output": "further cellular uptake analysis", "reagent": ["PBS"]}
Incubate HeLa cells with fluorescence-modified SPMNPs at 37°C for different Pulse–Chase periods.
{"action": "incubate", "output": "", "reagent": ["fluorescence-modified SPMNPs"], "time": ["different Pulse\u2013Chase periods"], "temperature": ["37°C"]}
After the Pulse-Chase paradigm, fix the cells in 4% paraformaldehyde on a coverslip with polymount with DAPI nucleus staining. #####%%%%% output "DAPI nucleus staining" are not used
{"action": "fix", "output": "DAPI nucleus staining", "reagent": ["4% paraformaldehyde"]}
Capture the images on a Zeiss Radiance 2100 confocal connected to an upright Nikon Eclipse E800 microscope using an oil immersion plan Apo 40xA/1.40 numerical aperture objective lens.
{"action": "collect", "output": "", "device": ["an upright Nikon Eclipse E800 microscope"]}
Do final processing using Adobe Photoshop (Adobe, CA) .
{"action": "process", "output": ""}
restrict to limited linear color balance adjustments to interpret merged pictures
