---------- Forwarded message ---------
From: Ariel Zeleznikow-Johnston from Preserving Hope <
[email protected]>
Date: Thu, Jul 3, 2025 at 7:26 AM
Subject: Setting standards for brain preservation quality
To: <[email protected]>


Monitoring preservation’s transition from the laboratory into clinical
practice
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Setting standards for brain preservation quality
<https://substack.com/app-link/post?publication_id=2285419&post_id=166707345&utm_source=post-email-title&utm_campaign=email-post-title&isFreemail=true&r=6x3nn&token=eyJ1c2VyX2lkIjoxMTYyMjA4MywicG9zdF9pZCI6MTY2NzA3MzQ1LCJpYXQiOjE3NTE1NDE5NzIsImV4cCI6MTc1NDEzMzk3MiwiaXNzIjoicHViLTIyODU0MTkiLCJzdWIiOiJwb3N0LXJlYWN0aW9uIn0.c5m0jVQSeSuviznt9R-d3m6xheQYjwOmcxxLS14knnk>Monitoring
preservation’s transition from the laboratory into clinical practice

Ariel Zeleznikow-Johnston <https://substack.com/@preservinghope>
Jul 3
<https://substack.com/@preservinghope>

<https://substack.com/app-link/post?publication_id=2285419&post_id=166707345&utm_source=substack&isFreemail=true&submitLike=true&token=eyJ1c2VyX2lkIjoxMTYyMjA4MywicG9zdF9pZCI6MTY2NzA3MzQ1LCJyZWFjdGlvbiI6IuKdpCIsImlhdCI6MTc1MTU0MTk3MiwiZXhwIjoxNzU0MTMzOTcyLCJpc3MiOiJwdWItMjI4NTQxOSIsInN1YiI6InJlYWN0aW9uIn0.Mp01iu_UQHBPAyAkHXptPv9B3vm3ePyhcahiNzu-dHc&utm_medium=email&utm_campaign=email-reaction&r=6x3nn>
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*This update has also been posted
<https://substack.com/redirect/186791e2-8e5a-4b79-bff9-caacc3af9539?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
on the Brain Preservation Foundation’s website*
Summary

The potential of preserving people for possible future revival depends on
maintaining the neural structures that encode memories, personality, and
other aspects of personal identity. Ensuring preservation procedures
actually achieve this requires rigorous assessment standards to verify that
critical brain structures remain intact. Historically, there have been no
standardized quality metrics or independent third-party evaluation of
preservation providers, making it difficult for patients to assess
different procedures or for the field to demonstrate scientific legitimacy.
To address this, the Brain Preservation Foundation intends to establish
quality standards and an accreditation program to fill this gap, providing
independent assessment of preservation providers to improve technical
rigor, encourage best practices, and help transition high-quality brain
preservation from laboratory research into credible clinical practice.
------------------------------
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Background

By preserving the brains and bodies of dying people, we may be able to
provide them a chance of eventual revival as medical technology advances
over the coming decades or centuries. The foundational premise of brain
preservation for life extension
<https://substack.com/redirect/5d16ed3c-1292-497b-9b99-b4d3e873684e?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
is that by preventing someone’s brain from decaying - by stabilising the
neural structures that underlie someone’s memories, personality, and other
psychological properties - the potential remains for this person to one day
be revived. As in cases of deep anaesthesia or induced hypothermia
<https://substack.com/redirect/415b25ee-1814-4b85-aa92-ba06c5eb6afe?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>,
where patients undergo complete (but reversible) cessation of brain
activity, so too may individuals be able to survive indefinite periods of
stasis, so long as their brains are preserved sufficiently well.

The viability of this approach fundamentally depends on preserving the
personal-identity-defining information content that underlies who someone
is. Current neuroscientific consensus
<https://substack.com/redirect/9480a397-ed9e-46fd-975e-e21fe5b17975?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
holds that information in human brains retained for anything longer than
short durations is primarily stored in the pattern of synaptic connections
between neurons. More specifically, the constancy of memories and
personality depend on maintaining specific connection strengths between
neurons, which in turn rely on neurophysiological properties such as
receptor types and densities, axonal myelination, perineuronal net
modifications, and other molecular and structural features.

Building on this scientific foundation, in 2010 the Brain Preservation
Foundation established a prize
<https://substack.com/redirect/eef189e1-44fd-40f4-ae9c-7fc6e980dcac?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
for the first team to rigorously demonstrate a technique capable of
preserving an entire brain for long-term storage with such fidelity that
“the structure of every neuronal process and every synaptic connection
remains intact and traceable using today's electron microscopic imaging
techniques.” While additional factors beyond traceability - such as
specific receptor types, intrinsic neuronal plasticity, and molecular
phosphorylation states - may also prove important for complete information
retention, ensuring comprehensive traceability of neuronal processes and
synaptic connections provides an evidence-based minimum standard for
preservation quality assessment. The prize was eventually won in 2018
<https://substack.com/redirect/ff900c8e-53d7-4ca3-ba12-e2a6ddcbdc7c?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
with the development of aldehyde-stabilized cryopreservation (ASC), with
rigorous assessment showing that it could meet this concrete benchmark for
preservation quality.
<https://substack.com/redirect/7ac81191-6164-4fa4-8ee0-be15856f0ba8?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
A)
FIBSEM electron micrograph taken of a section of pig cortex preserved using
aldehyde-stabilised cryopreservation. Taken as part of the BPF’s ASC
Evaluation images. B) Transmission electron micrograph of a section of a
cryopreserved rabbit brain. Provided by 21st Century Medicine.

In contrast, historical attempts to preserve patients for future revival
have not met this minimum standard. Traditional cryopreservation techniques
used on human patients have resulted in severe dehydration, causing brain
shrinkage of 30-50%
<https://substack.com/redirect/36fad7c7-6ea0-4904-83da-1406ca70ed5f?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>.
Animal experiments demonstrate that brain tissue preserved using these
methods appears highly compressed
<https://substack.com/redirect/da03ef6d-19bf-4a9b-a20d-7604ae72cecd?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
and structurally abnormal when viewed using powerful microscopes, such as
electron microscopy. While it remains theoretically conceivable that such
shrinkage might occur while preserving synaptic connectivity information,
it is equally plausible that this contraction masks osmotic and ischemic
damage, or other information-destroying alterations (e.g. torn axons or
displaced neural processes).

This unclear preservation quality presents a barrier for terminally ill
patients seeking preservation options. Without reliable quality assessment,
patients cannot easily distinguish the probability with which different
procedures offer some guarantee of preservation of their neural
information. Moreover, the lack of standardized quality metrics makes it
difficult for researchers and providers to improve their techniques, or for
the broader scientific and medical communities to evaluate the field's
legitimacy.

This is unfortunate, as recent advances in preservation technology -
particularly the development of ASC - have demonstrated that high-quality
whole-brain preservation is technically achievable. Now is the time for
translating these laboratory breakthroughs into clinical practice. However,
doing so requires robust quality standards and oversight mechanisms to
ensure consistent application and continuous improvement by preservation
providers.

The Brain Preservation Foundation's mission is to promote validated
scientific research and technical services development in the field of
whole brain preservation and to advocate for guidelines, policies, and
strategies that will increase the quality of brain preservation choice. By
developing comprehensive quality standards and an accreditation program, we
hope to enable high-quality preservation to now move beyond the laboratory
and into clinical practice.
Draft standards and accreditation processGeneral considerations

   1.

   These standards and accreditation processes are meant to be generic
   across all potential preservation protocols, whether they are fixative,
   cryopreservative, or as-yet undiscovered methods.
   2.

   These standards will almost certainly evolve over time, as preservation
   methods and readouts develop further and the basis of neurophysiological
   information storage becomes better understood.
   3.

   These standards are designed with the assumption that a patient’s brain
   was structurally intact and had suffered minimal damage or decay prior to
   the commencement of the preservation procedure. This assumption may often
   be violated in actual clinical settings, such as with patients who undergo
   prolonged agonal phases involving hypoxia and hypoperfusion, or who suffer
   from advanced dementia.

Standards

*Macroscopic*

   -

   *Structural integrity*: The brain must retain its overall anatomical
   structure with no gross fragmentation or tissue loss.
   -

   *Uniform preservation*: All brain regions must show consistent
   preservation quality with no areas of obvious deterioration.

*Microscopic*

   -

   *Neuronal process traceability*: Individual neural processes (i.e.
   axons, dendrites) must be continuously traceable without breaks or
   discontinuities.
   -

   *Membrane structure integrity*: Cell membranes, including synaptic
   membranes, must maintain their structural organization and be clearly
   distinguishable.
   -

   *Synapse identification & characterization: *Synapses should be clearly
   identifiable, and structural features known to be correlated with
   electrophysiological coupling strength (e.g. spine volume, postsynaptic
   density area) should be clearly visible.

Accreditation

*Phase 1: Protocol Submission and Review*
Preservation providers submit a comprehensive protocol for BPF evaluation.
This submission must include:

   -

   Detailed preservation procedures and techniques.
   -

   Critical steps and their plausible failure modes.
   -

   Quality assessment methods to be used for each case.
   -

   Non-destructive evaluation steps that will verify preservation quality.
   -

   Documentation of equipment, training, and facility requirements.

*Phase 2: Validation Testing*
Following protocol approval, providers must demonstrate their technique's
effectiveness through controlled testing using either:

   -

   Animal models (typically large mammals such as pigs), or
   -

   Human research donations where donors have specifically consented to
   destructive full-brain dissection for evaluation purposes.

This validation phase aims to prove that the non-destructive evaluation
methods proposed in the protocol are sufficient to reliably assess
preservation quality. Providers must demonstrate consistent results across
multiple test cases before being accredited.

*Phase 3: Patient Brain Preservation Monitoring*
Only after successful completion of validation testing may providers offer
preservation procedures to human patients. All accredited human cases must
be:

   -

   Pre-registered with the BPF prior to the procedure.
   -

      Note: this does not necessarily preclude a provider from performing
      preservations that are not pre-registered, such as in emergency cases.
      -

   Conducted according to the approved protocol.
   -

   Documented with complete quality evaluation results, including
   -

      At least one non-destructive method to assess perfusion quality of
      the brain with preservative chemicals, such as
      -

         CT perfusion
         
<https://substack.com/redirect/65a423bd-2bf9-412a-a9a6-d3f85358cf2e?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
         monitoring demonstrating perfusion of all brain regions with
preservative
         chemicals.

         
<https://substack.com/redirect/d6c13f46-6f36-4924-ac45-53af5e3db2ac?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
a)
         Transverse CT image of a healthy human brain during contrast
         administration. b) Time-concentration curves. c) Cerebral blood flow
         measured by CT perfusion during following contrast
administration. Images
         modified from Hoeffner et al., 2004.
         
<https://substack.com/redirect/65a423bd-2bf9-412a-a9a6-d3f85358cf2e?j=eyJ1IjoiNngzbm4ifQ.I1PMvYo4mI3PquTDRhL5Dev-9_ouIq3kw6ZhrVNsy8o>
         -

         X-ray Angiography
         -

         Perfusion MRI
         -

         Imaging methods which directly demonstrate chemical fixation or
         CPA equilibration of tissues
         -

      At least one minimally-invasive method to assess brain
      ultrastructure, such as a limited number of small punch biopsies analysed
      with
      -

         Electron microscopy
         -

         Expansion Microscopy

*Ongoing Oversight and Transparency*

Each pre-registered, anonymised case will be listed on a dedicated BPF
website and followed through to completion. All quality evaluations and
outcomes will be posted publicly, ensuring transparency and enabling
continuous improvement of preservation techniques. This public registry
will allow patients, families, and the broader scientific community to
assess provider performance and make informed decisions about preservation
options.

Should the evaluation of a pre-registered case show that it failed to meet
the standards, the BPF will publicly document the specific deficiencies,
and potentially either: 1) require successful completion of additional
validation testing before any future human cases can be pre-registered, or;
2) revoke the provider's accreditation status.

Note that while we seek to provide an accreditation service, it is
important to understand that the BPF will not have any legal or regulatory
means to regulate preservation providers directly. While we hope that
providers will engage with our accreditation process and seek to
self-adhere to our standards, we have no authority to compel them to do so,
nor to prevent them from withdrawing at a later time should they initially
engage.

We also note that this accreditation process will be set up in a
revenue-neutral manner, to ensure the BPF never has a financial incentive
to accredit providers or validate particular protocols or cases.
Conclusion

We hope that the development of rigorous preservation quality standards and
accreditation will represent a pivotal moment for the field of brain
preservation. If preservation providers are provided clear benchmarks, they
will be better positioned to offer procedures that meet neuroscientific
criteria for information retention. In contrast, without these frameworks,
promising research achievements like ASC risk remaining confined to
laboratory settings, while patients will continue to receive preservation
procedures of unverified quality. Additionally, establishing standards
demonstrates the field's commitment to empirical rigor and evidence-based
practice.

Every year, hundreds of thousands of terminally ill individuals face the
end of their lives with no viable medical options for survival. Many of
these individuals still have a strong will-to-live, and presumably would be
glad to receive more time if only an option were available. For these
patients, high-quality brain preservation currently provides their only
chance of living longer.

We invite preservation providers who are committed to meeting rigorous
scientific standards to engage with this accreditation process. Similarly,
we welcome researchers, clinicians, bioethicists, funders and policymakers
who recognize the importance of establishing proper oversight for this
emerging field to join us. For those interested in supporting this work -
whether through funding or technical expertise - we welcome your input.
Please reach out to us at: *[email protected]
<[email protected]>*

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