Figure 4

Microspore sorting. (A) Pollen population is characterized by an elevated high angle scatter (SSC) and autofluorescence (observed in the GFP channel using a 530/40 nm bandpass filter). Within this population, microspores (right panel, circular gate) can be differentiated from bicellular and tricellular pollen by their characteristic smaller size, captured by a diminished low angle scatter (FSC) and time-of-flight (Pulse Width), as compared to other stages of pollen development. (B) Sorted microspores were inspected by microscopy to show purity and integrity, as revealed by DAPI staining. Scale bars: 30 μm (left panel) and 10 μm (right panels). (C) Sorted microspores stained with DAPI were counted on a wide-field fluorescence microscope to confirm that most sorted cells are Uninucleate Microspores (UNM). BCP - Bicellular Pollen.