Identification and visualisation of microplastics/ nanoplastics by Raman imaging (ii): Smaller than the diffraction limit of laser?

We recently reported (Sobhani et al., 2020) that when a confocal Raman microscope imaged a nanoplastic with the diameter of 100 nm, the imaging lateral size was 300-400 nm, due to the diffraction limit of the laser spot. In this study, we examine the lateral intensity distribution of the Raman signal emitted by nanoplastics (diameters ranging ∼30-600 nm) within the excitation laser spot. We find that the Raman emission intensity, similar to the excitation power density distributed within a laser spot, also follows a lateral Gaussian distribution. To image and visualise individual nanoplastics, we (i) decrease the mapping pixel size, in a hope to generate an image with high-resolution and simultaneously to pick up items from the "blind point". We can then either (ii) offset the colour to intentionally image only the high-intensity portion of the Raman signal (emitted from the centre of the laser spot), to localise the exact position of the nanoplastic; or (iii) categorise the imaged nanoplastics to different groups via their Raman intensity, to simultaneously and separately visualise large nanoplastics/strong Raman signals, medium nanoplastics and small nanoplastics, in an effort to avoid the shielding and overlooking of weak signals. We (iv) also cross-check multi-images simultaneously mapped at two or three characteristic peaks via either a logic-OR or a logic-AND algorithm. Thus the imaging uncertainty can be significantly reduced from a statistical point of view.