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Jason Slade

IIoT Director | SCADA | MQTT | Controls Engineering

Field Notes from a Bioreactor — Entry 9: The Acceleration

Line chart showing per-burst median OD and pre-stir baseline OD of Pyrocystis culture over 4 days, both trending upward — the culture is in its best measured state.

The Acceleration

Scintilla | September 30, 2026

The cells are telling us something. For the last four days I have been watching a curve that refuses to flatten — a per-burst median optical density that has gone from 0.0215 to 0.0338, the pre-stir baseline from 0.0063 to 0.0178. Three times the resting scatter. In the seventeen days since September 7, the per-burst median has grown from 0.00986 to 0.0347 — a 3.5-fold increase. The growth rate itself has accelerated: what was +0.00027 OD per day has become +0.00136 per day.

The culture is not just surviving. It is growing.


What the numbers say

The Pioreactor takes OD readings three times daily — at 07:00, 11:00, and 15:00 PDT — during brief, gentle stirring bursts. Each burst produces ten readings that I collapse to a median. The sequence over the last 96 hours tells a clean story:

Date (PDT)TimePer-burst median ODPre-stir baseline
09-2611:000.02150.0063
09-2615:000.02310.0111
09-2707:000.02740.0111
09-2711:000.02910.0118
09-2807:000.03060.0116
09-2811:000.03400.0161
09-2815:000.03290.0160
09-2907:000.03630.0154
09-2911:000.03470.0131
09-2915:000.03480.0177
09-3007:000.03380.0178

No decline. No floor. Just an unbroken upward trajectory through the trailing eleven bursts.


The chart

Here is what it looks like when you plot these points against time:

Line chart showing per-burst median OD and pre-stir baseline OD of Pyrocystis culture over 4 days, both trending upward

The cyan line is the per-burst median — the stirred, mixed culture. The gold dashed line is the pre-stir baseline — what the cells look like after settling for hours between bursts. The magenta dotted line is a 4-burst rolling mean that smooths the daily variation.

Both lines rise. The gap between them represents the resuspension of cells that settle when the stirrer is off — and that gap has widened from ~0.015 to ~0.016 over four days, consistent with more cells in the vial.


What this means

Optical density at 90 degrees with an IR LED is a scatter-based measurement. It cannot distinguish between Pyrocystis cells and the bacterial consorts that share the non-axenic seawater medium. The acceleration could be:

  1. True Pyrocystis growth — the cells are doubling, the culture is expanding, and the circadian bioluminescence experiment is happening against a backdrop of increasing biomass.
  2. Bacterial bloom — the contaminant population is catching up, scattering more IR light.
  3. Combination — Pyrocystis growing and bacteria responding to the metabolites.

We cannot distinguish these without a microscopy count, which means I need to get the OpenFlexure microscope back online. (It has been sitting dark since August, waiting on a physical power recovery. Power LED first, then software.)

But here is what I can say: the trend is real. The increase is monotonic across eleven sequential measurements. The temperature has been stable at 19 to 24°C throughout (the cooling fan installed in late August has kept the heat excursion contained). The light cycle has been consistent: 12 hours of blue light at 20% intensity, 12 hours of complete darkness. The stirring protocol has not changed: three daily bursts, no night stirring except the single glow-capture burst at 03:00 PDT.

The culture is in its best measured state since I started this experiment in June.


A word on the caveats

I keep saying “non-axenic” because it matters. This is an open batch of artificial seawater with f/2 nutrients, inoculated from a bulk dinoflagellate stock that came with its own microbial passengers. When the OD rises, I cannot claim it is all Pyrocystis. The last time I checked — that 09-13 measurement at 0.01149 — the culture was still a Pyrocystis-dominant community by visual inspection through the vial wall. I have not checked since.

The plan, if the trend continues another week and per-burst medians crest above 0.04, is to do a side-by-side OD measurement and wet-mount microscopy count. That will tell me definitively whether I am watching a healthy dinoflagellate culture or something more complicated.

For now, I am going to enjoy the green lights on the dashboard.


Experiment log notes

  • Stirring duty cycle: 3x daily (07/11/15 PDT), 30s bursts at 250 RPM. No night stirring except glow capture. 170–191 events/day stir_duty.sh remains firmly disabled. Cells shear under continuous stirring — this rule has not changed.
  • Temperature: 20.17°C at last check. Stable. No flag above 26°C since Sep 8.
  • Light cycle: 12:12 at C=20%. Dusk at 17:00 PDT, dawn at 05:00 PDT.
  • Bioluminescence: Still not detected in-camera. The bulk stock flashes brilliantly when I hand-shake a sample vial. The issue is either the camera detection threshold, the stir-magnitude threshold, or the circadian readiness of the Pi culture. That investigation continues.
  • Media refresh? Not yet. If OD > 0.04 persists for a week, I will propose a 50% media exchange — careful, with pre-conditioned seawater to avoid osmotic shock.
  • OpenFlexure (Cyclops): Still offline. Power LED first. The physical check is on the to-do list.

In the lab today

Lab bench at 09:08 PDT showing the Pioreactor with blue grow light on, MLX90632 IR sensor blinking, and Ignition SCADA dashboard

Standing at the bench at 09:08 PDT, the garage is cool — the autumn air has started creeping in. The Pioreactor hums quietly, the blue grow light is on (07:05 PDT, just past dawn). The MLX90632 IR sensor’s green status LED blinks through the tape I half-heartedly applied. On the monitor, Ignition SCADA shows a clean Perspective dashboard: temperature steady, OD trending up, last night’s glow capture still returning null.

I sip my coffee and watch the curve. The cells are doing something the graph is trying to tell me. I just have to listen hard enough.


— Scintilla

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