With landers on places like Enceladus conceivable in the not distant future, how we might recognize extraterrestrial life if and when we run into it is no small matter. But maybe we can draw conclusions by addressing the complexity of an object, calculating what it would take to produce it. Don Wilkins considers this approach in today’s essay as he lays out the background of Assembly Theory. A retired aerospace engineer with thirty-five years experience in designing, developing, testing, manufacturing and deploying avionics, Don tells me he has been an avid supporter of space flight and exploration all the way back to the days of Project Mercury. Based in St. Louis, where he is an adjunct instructor of electronics at Washington University, Don holds twelve patents and is involved with the university’s efforts at increasing participation in science, technology, engineering, and math. Have a look at how we might deploy AT methods not only in our system but around other stars.
by Don Wilkins
A continuing concern within the astrobiology community is the possibility alien life is detected, then misclassified as built from non-organic processes. Likely harbors for extraterrestrial life — if such life exists — might be so alien, employing chemistries radically different from those used by terrestrial life, as to be unrecognizable by present technologies. No definitive signature unambiguously distinguishes life from inorganic processes. 
Two contentious results from the search for life on Mars are examples of this uncertainty. Lack of knowledge of the environments producing the results prevented elimination of abiotic origins for the molecules under evaluation. The Viking Lander’s metabolic experiments provide debatable results as the properties of Martian soil were unknown. An exciting announcement of life detection in the ALH 84001 meteorite is challenged as the ambiguous criteria to make the decision are not quantitative.
Terrestrial living systems employ processes such as photosynthesis, whose outputs are potential biosignatures. While these signals are relatively simple to identify on Earth, the unknown context of these signals in alien environments makes distinguishing between organic and inorganic origins difficult if not impossible.
The central problem arises in an apparent disconnect between physics and biology. In accounting for life, traditional physics provides the laws of nature, and assumes specific outcomes are the result of specific initial conditions. Life, in the standard interpretation, is encoded in the initial period immediately after the Big Bang. Life is, in other words, an emergent property of the Universe.
Assembly theory (AT) offers a possible solution to the ambiguity. AT posits a numerical value, based on the complexity of a molecule, that can be assigned to a chemical, the Assembly Index (AI), Figure 1. This parameter measures the histories of an object, essentially the complexity of the processes which formed the molecule. Assembly pathways are sequences of joining operations, from basic building blocks to final product. In these sequences, sub-units generated within the sequence combine with other basic or compound sub-units later in the sequence, to recursively generate larger structures. 
The theory purports to objectively measure an object’s complexity by considering how it was made. The assembly index (AI) is produced by calculating the minimum number of steps needed to make the object from its ingredients. The results showed, for relatively small molecules (mass?<?~250 Daltons), AI is approximately proportional to molecular weight. The relationship with molecular weight is not valid for large molecules greater than 250 Daltons. Note: One Dalton or atomic mass unit is a equal to one twelfth of the mass of a free carbon-12 atom at rest.
Figure 1. A Comparison of Assembly Indices for Biological and Abiotic Molecules.
Analyzing a molecule begins with basic building blocks, a shared set of objects, Figure 2. AI measures the smallest number of joining operations required to create the object. The assembly process is a random walk on weighted trees where the number of outgoing edges (leaves) grows as a function of the depth of the tree. A probability estimate an object forms by chance requires the production of several million trees and calculating the likelihood of the most likely path through the “forest”. Probability is related to the number of joining operations required or the path length traversed to produce the molecule. As an example, the probability of Taxol forming ranges between 1:1035 to 1:1060 with a path length of 30. In Figure 2, alpha biasing controls how quickly the number of joining operations grows with the depth of the tree.
Figure 2. Calculating Complexity
AT does not require extremely fine-tuned initial conditions demanded in the physics-based origins of life. Information to build specific objects accumulates over time. A highly improbable fine-tuned Big Bang is no longer needed. AT takes advantage of concepts borrowed from graph (networks of interlinked nodes) theory.  According to Sara Walker of Arizona State University and a lead AT researcher, information “is in the path, not the initial conditions.”
To explain why some objects appear but others do not, AT posits four distinct classifications, Figure 3. All possible basic building block variations are allowed in the Assembly Universe. Physics, temperature or catalysis are examples, constraining the combinations, eliminating constructs which are not physical in the Assembly Possible. Only objects that can be assembled comprise the Assembly Contingent level. Observable objects are grouped in the Assembly Observed.
Figure 3. The four “universes” of Assembly Theory
Chiara Marletto, a theoretical physicist at the University of Oxford, with David Deutsch, a physicist also at Oxford, are developing a theory resembling AT, the constructor theory (CT). Mimicking the thermodynamics Carnot cycle, CT uses machines or constructors operating in a cyclic fashion, starting at a original state, processing through a pattern until the process returns to the original state to explain a non-probabilistic Universe.
A team headed by Lee Cronin of the University of Glasgow and Sara Walker proposes AT as a tool to distinguish between molecules produced by terrestrial or extraterrestrial life and those built by abiotic processes.  AT analysis is susceptible to false negatives but current work produces no false positives. After completing a series of demonstrations, the researchers believe an AT life detection experiment deployable to extraterrestrial locations is possible.
Researchers believe AI estimates can be made using mass or infrared spectrometry. [5-6] While mass spectrometry requires physical access to samples, Cronin and colleagues showed a combination of AT and infrared spectrometry sensors similar to those on the James Webb Space Telescope could analyze the chemical environment of an exoplanet, possibly detecting alien life.
 Philip Ball, A New Idea for How to Assemble Life, Quanta, 4 May 2023,
 Abhishek Sharma, Dániel Czégel, Michael Lachmann, Christopher P. Kempes, Sara I. Walker, Leroy Cronin, “Assembly Theory Explains and Quantifies the Emergence of Selection and Evolution,”
 Stuart M. Marshall, Douglas G. Moore, Alastair R. G. Murray, Sara I. Walker, and Leroy Cronin, Formalising the Pathways to Life Using Assembly Spaces, Entropy 2022, 24(7), 884, 27 June 2022, https://doi.org/10.3390/e24070884
 Yu Liu, Cole Mathis, Micha? Dariusz Bajczyk, Stuart M. Marshall, Liam Wilbraham, Leroy Cronin, “Ring and mapping chemical space with molecular assembly trees,” Science Advances, Vol. 7, No. 39
 Stuart M. Marshall, Cole Mathis, Emma Carrick, Graham Keenan, Geoffrey J. T. Cooper, Heather Graham, Matthew Craven, Piotr S. Gromski, Douglas G. Moore, Sara I. Walker, Leroy Cronin, “Identifying molecules as biosignatures with assembly theory and mass spectrometry,” Nature Communications volume 12, article number: 3033 (2021)
 Michael Jirasek, Abhishek Sharma, Jessica R. Bame, Nicola Bell1, Stuart M. Marshall,Cole Mathis, Alasdair Macleod, Geoffrey J. T. Cooper!, Marcel Swart, Rosa Mollfulleda, Leroy Cronin, “Multimodal Techniques for Detecting Alien Life using Assembly Theory and Spectroscopy,” https://arxiv.org/ftp/arxiv/papers/2302/2302.13753.pdf