A Molecular Logic Gate Enables Unconventional Super-resolution Same-Day Imaging of Lysosome Membrane in Live Cells

Lysosomes are acidic membrane-bound organelles that aid digestion, excretion, and cell renewal. The lysosomal membranes are essential for maintaining lysosomal functions and cellular homeostasis. In this work, we developed a molecular “NOR” logic gate, SIATFluor-580L, by introducing malachite green into the spirocyclic rhodamine. SIATFluor-580L requires restriction of molecular rotation of the malachite green motif (Input 1, tight membrane structure) and a large amount of H⁺ ions to convert the spirocyclic rhodamine into the zwitterionic form (Input 2, acidic environment) to produce a fluorescent product (Output), providing a fluorogenic probe to visualize the lysosomal membrane dynamics in living cells with subdiffraction resolution by using HyVolution (also known as Lightning), an unconventional and inexpensive super-resolution imaging approach based on a basic confocal optical system.