HOW IT WORKS
Fluorescence Membrane Microscopy: What You Need to Know
Fluorescence Membrane Microscopy (FMM) works with Backgrounded Membrane Imaging (BMI) on the Aura to tell you exactly what is in your formulation. It utilizes fluorescent reagents that bind specifically to a particular particle population greater than 5µm in size for simple and definitive identification that doesn’t depend on potentially misleading morphology and intensity data.
Membrane phase stain and solution phase staining are both compatible with Aura, so you can choose whichever method works best for your workflow. To membrane phase stain, just perform the normal steps required for BMI. Then, add 40 µL of fluorescent reagent to the plate, apply vacuum, and then take your FMM measurement. Solution phase staining is just as simple, you just mix the reagent fluorescent dye with the sample in a test tube before applying it to a backgrounded plate and taking your BMI and FMM measurements. When you’re done, our software will automatically align all the images generated so you can jump right into your analysis.
Red – protein aggregates
Black – non-protein particles
Green – degraded polysorbate
Blue – side illumination mode (SIMI)
Excitation | Emission | Example Dye | |
---|---|---|---|
Channel 1 | 440/40 nm | 500/40 nm | Thioflavin T |
Channel 2 | 376/30 nm 482/35 488/50 540/50 605/50 | 440/40 nm 524/24 544/24 600/37 670/50 | TMP-DPH, DAPI BODIPY FL C16 SYBR™ Gold Dil |
Channel 3 (Aura+ only) | 376/30 nm 48235 488/50 540/50 605/50 | 440/40 nm 524/24 544/24 600/37 670/50 | TMP-DPH, DAPI BODIPY FL C16 SYBR™ Gold Dil |
Custom (Aura only) | Custom | Custom | Dye of choice |