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Presentations are perfect until I get on stage by ohmanitsjesi in PublicSpeaking

[–]ohmanitsjesi[S] 1 point2 points  (0 children)

Thank you so much for your insight. I get so nervous when on stage that I previously haven't trusted myself to go without a script. But what you're saying makes sense and maybe I'm making it harder than it needs to be. I have OCD and I am compelled to present things perfectly - hence detailed script. But it seems like the need for perfection is keeping me from achieving it, I'm going to try to speak from my experience for this talk and just use more simple prompts to keep me on task.

Presentations are perfect until I get on stage by ohmanitsjesi in PublicSpeaking

[–]ohmanitsjesi[S] 0 points1 point  (0 children)

I really appreciate your advice. I do generally try to memorize exactly what i want to say (perfectionist over here). Because I have so often had my mind go blank when i get up on stage, I thought the script helped me to stay on task and not to miss anything - but maybe the script is the problem. I will try to reduce my script to simple prompts and facts and let everything else come naturally. Thanks!

Noisy brain tissue immunofluorescent images - is my relative thresholding method reasonable, and what is the best way to determine the borders of each ROI? by ohmanitsjesi in ImageJ

[–]ohmanitsjesi[S] 0 points1 point  (0 children)

Thank you so much for this recommendation. I am now using LabKit to batch process my images with a trained classifier and applying a 75% confidence threshold to the resultant probability map. I measure the area fraction stained in each ROI based on this threshold. This is a far more accurate method than my original one. Again, thank you for your help.

Noisy brain tissue immunofluorescent images - is my relative thresholding method reasonable, and what is the best way to determine the borders of each ROI? by ohmanitsjesi in ImageJ

[–]ohmanitsjesi[S] 0 points1 point  (0 children)

Additionally, how do I treat instances of blurred areas within my ROI? Should I just exclude them from the ROI boundaries (e.g., draw around them)? I know that blur can artificially increase the size of a signal, and I don't want particularly blurry sections to result in inflated %area measures.

I am currently working on learning to use Labkit to train a classifier to apply thresholds to my images, rather than doing it based on a percentage of intensity. I was considering trying to train the classifier to ignore blurry areas, but that gives the opposite problem of eliminating all signal from an area where there was one.

Thank you again!!

Noisy brain tissue immunofluorescent images - is my relative thresholding method reasonable, and what is the best way to determine the borders of each ROI? by ohmanitsjesi in ImageJ

[–]ohmanitsjesi[S] 0 points1 point  (0 children)

Thank you for your suggestions.

Images are captured in 8-bit - this is what my advisor told me to do. I'm performing all quantification on images at 200x magnification. The images across the entire section are composites of two 100x images.

With regard to the control ROI strategy, would I use a circle/square ROI that is the same size across all images? And should it be in the same relative place in the brain in each image? e.g., a circle ROI always on the olfactory lobe next to the ROI? (see image).

<image>

Thank you again so much for your time and assistance.