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reddot2  (Biotium)
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Experimental overview to measure fluorescent Alexa-488 labeled molecules’ nuclear uptake in cells. (a) Graphical representation of experimental setup. (b) Example of an individual nucleus’ intensity increase in the cargo channel. Raw data are circular points while the blue line is a polynomial best fit used to extract the, while the maximum rate of nuclear uptake was found by obtaining the derivative of the best fit line along every point of the curve. The dashed black line is the or half of the total uptake of the molecule for that given nucleus, whereas the dotted gray vertical line is the , or the time at which half of the uptake has occurred; both are used to obtain population statistics displayed in later figures. The y-axis on this graph is based on the post-processing software CellProfiler that use a 0–1 re-scaled fluorescent counts based on the maximum possible number associated with the bit count (for a 16 bit image it scales of the maximum of 65,535). The experimental time extends past where the graph cuts off. Here, we focus on early time points to graphically emphasize the quantities used for statistical comparison. (c) Exemplary images of the expierment at different times. Time 0 shows the <t>RedDot2</t> channel that highlights (bright) nuclei that will be used as ROIs. This is before and cargo has been added. The three green colored images are of the Alexa-488 conjugated 10 kDa dextran channel at the indicated times. Passive nuclear uptake is seen in permeabilized cells at longer times (marked by dark pink arrows). Dark cells in the later time points (marked by light pink arrows) correspond to non-permeabilized cells. Figure compiled in BioRender.
Reddot2, supplied by Biotium, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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reddot 2  (Biotium)
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Biotium reddot 2
Experimental overview to measure fluorescent Alexa-488 labeled molecules’ nuclear uptake in cells. (a) Graphical representation of experimental setup. (b) Example of an individual nucleus’ intensity increase in the cargo channel. Raw data are circular points while the blue line is a polynomial best fit used to extract the, while the maximum rate of nuclear uptake was found by obtaining the derivative of the best fit line along every point of the curve. The dashed black line is the or half of the total uptake of the molecule for that given nucleus, whereas the dotted gray vertical line is the , or the time at which half of the uptake has occurred; both are used to obtain population statistics displayed in later figures. The y-axis on this graph is based on the post-processing software CellProfiler that use a 0–1 re-scaled fluorescent counts based on the maximum possible number associated with the bit count (for a 16 bit image it scales of the maximum of 65,535). The experimental time extends past where the graph cuts off. Here, we focus on early time points to graphically emphasize the quantities used for statistical comparison. (c) Exemplary images of the expierment at different times. Time 0 shows the <t>RedDot2</t> channel that highlights (bright) nuclei that will be used as ROIs. This is before and cargo has been added. The three green colored images are of the Alexa-488 conjugated 10 kDa dextran channel at the indicated times. Passive nuclear uptake is seen in permeabilized cells at longer times (marked by dark pink arrows). Dark cells in the later time points (marked by light pink arrows) correspond to non-permeabilized cells. Figure compiled in BioRender.
Reddot 2, supplied by Biotium, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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nuclear stain  (Biotium)
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Experimental overview to measure fluorescent Alexa-488 labeled molecules’ nuclear uptake in cells. (a) Graphical representation of experimental setup. (b) Example of an individual nucleus’ intensity increase in the cargo channel. Raw data are circular points while the blue line is a polynomial best fit used to extract the, while the maximum rate of nuclear uptake was found by obtaining the derivative of the best fit line along every point of the curve. The dashed black line is the or half of the total uptake of the molecule for that given nucleus, whereas the dotted gray vertical line is the , or the time at which half of the uptake has occurred; both are used to obtain population statistics displayed in later figures. The y-axis on this graph is based on the post-processing software CellProfiler that use a 0–1 re-scaled fluorescent counts based on the maximum possible number associated with the bit count (for a 16 bit image it scales of the maximum of 65,535). The experimental time extends past where the graph cuts off. Here, we focus on early time points to graphically emphasize the quantities used for statistical comparison. (c) Exemplary images of the expierment at different times. Time 0 shows the <t>RedDot2</t> channel that highlights (bright) nuclei that will be used as ROIs. This is before and cargo has been added. The three green colored images are of the Alexa-488 conjugated 10 kDa dextran channel at the indicated times. Passive nuclear uptake is seen in permeabilized cells at longer times (marked by dark pink arrows). Dark cells in the later time points (marked by light pink arrows) correspond to non-permeabilized cells. Figure compiled in BioRender.
Nuclear Stain, supplied by Biotium, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Experimental overview to measure fluorescent Alexa-488 labeled molecules’ nuclear uptake in cells. (a) Graphical representation of experimental setup. (b) Example of an individual nucleus’ intensity increase in the cargo channel. Raw data are circular points while the blue line is a polynomial best fit used to extract the, while the maximum rate of nuclear uptake was found by obtaining the derivative of the best fit line along every point of the curve. The dashed black line is the or half of the total uptake of the molecule for that given nucleus, whereas the dotted gray vertical line is the , or the time at which half of the uptake has occurred; both are used to obtain population statistics displayed in later figures. The y-axis on this graph is based on the post-processing software CellProfiler that use a 0–1 re-scaled fluorescent counts based on the maximum possible number associated with the bit count (for a 16 bit image it scales of the maximum of 65,535). The experimental time extends past where the graph cuts off. Here, we focus on early time points to graphically emphasize the quantities used for statistical comparison. (c) Exemplary images of the expierment at different times. Time 0 shows the RedDot2 channel that highlights (bright) nuclei that will be used as ROIs. This is before and cargo has been added. The three green colored images are of the Alexa-488 conjugated 10 kDa dextran channel at the indicated times. Passive nuclear uptake is seen in permeabilized cells at longer times (marked by dark pink arrows). Dark cells in the later time points (marked by light pink arrows) correspond to non-permeabilized cells. Figure compiled in BioRender.

Journal: Nucleus

Article Title: Passive nuclear transport deviates from Fickian behavior in prostate and breast cell types

doi: 10.1080/19491034.2026.2620223

Figure Lengend Snippet: Experimental overview to measure fluorescent Alexa-488 labeled molecules’ nuclear uptake in cells. (a) Graphical representation of experimental setup. (b) Example of an individual nucleus’ intensity increase in the cargo channel. Raw data are circular points while the blue line is a polynomial best fit used to extract the, while the maximum rate of nuclear uptake was found by obtaining the derivative of the best fit line along every point of the curve. The dashed black line is the or half of the total uptake of the molecule for that given nucleus, whereas the dotted gray vertical line is the , or the time at which half of the uptake has occurred; both are used to obtain population statistics displayed in later figures. The y-axis on this graph is based on the post-processing software CellProfiler that use a 0–1 re-scaled fluorescent counts based on the maximum possible number associated with the bit count (for a 16 bit image it scales of the maximum of 65,535). The experimental time extends past where the graph cuts off. Here, we focus on early time points to graphically emphasize the quantities used for statistical comparison. (c) Exemplary images of the expierment at different times. Time 0 shows the RedDot2 channel that highlights (bright) nuclei that will be used as ROIs. This is before and cargo has been added. The three green colored images are of the Alexa-488 conjugated 10 kDa dextran channel at the indicated times. Passive nuclear uptake is seen in permeabilized cells at longer times (marked by dark pink arrows). Dark cells in the later time points (marked by light pink arrows) correspond to non-permeabilized cells. Figure compiled in BioRender.

Article Snippet: To stain the nuclei, RedDot2 (Biotium catalog # 40,061) with a dilution ratio of 1:200 should be prepared prior to experimentation within the digitonin solution.

Techniques: Labeling, Software, Comparison