Light Transmission Spectroscopy (LTS) is a recently developed technique for measuring the diameter and concentration of nanoparticles in fluid suspension. LTS is based on obtaining the optical extinction of light as a function of wavelength, over the range of ∼220–1100 nm, and using Mie theory to transform extinction information into a particle‐size distribution. This has been made practical with recent developments in compact, high resolution optical spectrometers and “lap‐top” computing. LTS has a particle diameter measurement range of ∼1–3000 nm. LTS has a large dynamic range, with an ultimate concentration range covering several orders of magnitude. The technique is most applicable to relatively dilute samples, with a minimum detection concentration of ∼1000 objects/mL. LTS has evolved from a laser‐based laboratory instrument to a transportable format with a modest bench‐top footprint. The system has proven itself especially useful to biological‐type investigations involving objects ranging in size from proteins to bacteria – and has even been adapted to detect species‐specific environmental DNA. A special feature of the technique is its ability to perform optical subtraction, whereby objects of a given size and concentration in a reference channel can effectively eliminate their optical contribution in a sample‐measurement channel.