Pulse–Chase with ¹⁴CO₂ — following a single carbon through the Calvin cycle

Every carbon atom here is a persistent object, so a labelled carbon can be followed from CO₂ into 3-PGA, around the regeneration shuffle and out as sucrose — including which position it occupies in each molecule. This recreates the experiment Calvin, Benson and Bassham used to deduce the cycle: label briefly, kill the sample at a chosen moment, separate the extract by two-dimensional paper chromatography, and read the radioactive spots off the film.

Stroma — pathway map

Node fill shows the fraction of that pool's carbon which is radioactive. Watch the label enter at RuBisCO, appear in 3-PGA first, then spread.

Label distribution over chase time

Each line is that compound's share of all label currently in the cycle. 3-PGA spikes first and then falls as the label moves downstream — the observation that identified 3-PGA as the first stable product of carbon fixation.

Experiment controls

Chase
0 / 0
¹⁴C in cycle
0
CO₂ fixed
0
Hexose out

Autoradiogram SAMPLE KILLED

Simulated 2-D paper chromatogram; darker spots carry more ¹⁴C. Spot positions are schematic — arranged by relative mobility (bisphosphates barely leave the origin, free sucrose runs furthest) rather than measured Rf values.

Where the label sits inside hexose

no label in hexose yet

Carbons 3 and 4 light up first. Both descend from the carboxyl carbon of 3-PGA — the position the incoming CO₂ becomes — which is exactly what aldolase's condensation of two triose phosphates predicts.

Follow one carbon